WorldWideScience

Sample records for nef reveals critical

  1. Tracking the Emergence of Host-Specific Simian Immunodeficiency Virus env and nef Populations Reveals nef Early Adaptation and Convergent Evolution in Brain of Naturally Progressing Rhesus Macaques.

    Science.gov (United States)

    Lamers, Susanna L; Nolan, David J; Rife, Brittany D; Fogel, Gary B; McGrath, Michael S; Burdo, Tricia H; Autissier, Patrick; Williams, Kenneth C; Goodenow, Maureen M; Salemi, Marco

    2015-08-01

    evolutionary patterns in the gp120 and nef genes leading to the emergence of host-specific viral populations and potentially linked to disease progression. Although each macaque exhibited unique immune profiles, macaque-specific nef sequences evolving under selection were consistently detected in plasma samples at 3 months postinfection, significantly earlier than in gp120 macaque-specific sequences. On the other hand, nef sequences in brain tissues, collected at necropsy of two animals with detectable infection in the central nervous system (CNS), revealed convergent evolution. The results not only indicate that early adaptation of nef in the new host may be essential for successful infection, but also suggest that specific nef variants may be required for SIV to efficiently invade CNS macrophages and/or enhance macrophage migration, resulting in HIV neuropathology. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  2. Proteomic analysis of HIV-1 Nef cellular binding partners reveals a role for exocyst complex proteins in mediating enhancement of intercellular nanotube formation

    Directory of Open Access Journals (Sweden)

    Mukerji Joya

    2012-06-01

    Full Text Available Abstract Background HIV-1 Nef protein contributes to pathogenesis via multiple functions that include enhancement of viral replication and infectivity, alteration of intracellular trafficking, and modulation of cellular signaling pathways. Nef stimulates formation of tunneling nanotubes and virological synapses, and is transferred to bystander cells via these intercellular contacts and secreted microvesicles. Nef associates with and activates Pak2, a kinase that regulates T-cell signaling and actin cytoskeleton dynamics, but how Nef promotes nanotube formation is unknown. Results To identify Nef binding partners involved in Pak2-association dependent Nef functions, we employed tandem mass spectrometry analysis of Nef immunocomplexes from Jurkat cells expressing wild-type Nef or Nef mutants defective for the ability to associate with Pak2 (F85L, F89H, H191F and A72P, A75P in NL4-3. We report that wild-type, but not mutant Nef, was associated with 5 components of the exocyst complex (EXOC1, EXOC2, EXOC3, EXOC4, and EXOC6, an octameric complex that tethers vesicles at the plasma membrane, regulates polarized exocytosis, and recruits membranes and proteins required for nanotube formation. Additionally, Pak2 kinase was associated exclusively with wild-type Nef. Association of EXOC1, EXOC2, EXOC3, and EXOC4 with wild-type, but not mutant Nef, was verified by co-immunoprecipitation assays in Jurkat cells. Furthermore, shRNA-mediated depletion of EXOC2 in Jurkat cells abrogated Nef-mediated enhancement of nanotube formation. Using bioinformatic tools, we visualized protein interaction networks that reveal functional linkages between Nef, the exocyst complex, and the cellular endocytic and exocytic trafficking machinery. Conclusions Exocyst complex proteins are likely a key effector of Nef-mediated enhancement of nanotube formation, and possibly microvesicle secretion. Linkages revealed between Nef and the exocyst complex suggest a new paradigm of

  3. HIV-1 Nef binds with human GCC185 protein and regulates mannose 6 phosphate receptor recycling

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, Manjeet; Kaur, Supinder; Nazir, Aamir; Tripathi, Raj Kamal, E-mail: rajkamalcdri@gmail.com

    2016-05-20

    HIV-1 Nef modulates cellular function that enhances viral replication in vivo which culminate into AIDS pathogenesis. With no enzymatic activity, Nef regulates cellular function through host protein interaction. Interestingly, trans-cellular introduction of recombinant Nef protein in Caenorhabditis elegans results in AIDS like pathogenesis which might share common pathophysiology because the gene sequence of C. elegans and humans share considerable homology. Therefore employing C. elegans based initial screen complemented with sequence based homology search we identified GCC185 as novel host protein interacting with HIV-1 Nef. The detailed molecular characterization revealed N-terminal EEEE{sub 65} acidic domain of Nef as key region for interaction. GCC185 is a tethering protein that binds with Rab9 transport vesicles. Our results show that Nef-GCC185 interaction disrupts Rab9 interaction resulting in delocalization of CI-MPR (cation independent Mannose 6 phosphate receptor) resulting in elevated secretion of hexosaminidase. In agreement with this, our studies identified novel host GCC185 protein that interacts with Nef EEEE65 acidic domain interfering GCC185-Rab9 vesicle membrane fusion responsible for retrograde vesicular transport of CI-MPR from late endosomes to TGN. In light of existing report suggesting critical role of Nef-GCC185 interaction reveals valuable mechanistic insights affecting specific protein transport pathway in docking of late endosome derived Rab9 bearing transport vesicle at TGN elucidating role of Nef during viral pathogenesis. -- Highlights: •Nef, an accessory protein of HIV-1 interacts with host factor and culminates into AIDS pathogenesis. •Using Caenorhabditis elegans based screen system, novel Nef interacting cellular protein GCC185 was identified. •Molecular characterization of Nef and human protein GCC185 revealed Nef EEEE{sub 65} key region interacted with full length GCC185. •Nef impeded the GCC185-Rab 9 interaction and

  4. Proteomic profiling of SupT1 cells reveal modulation of host proteins by HIV-1 Nef variants.

    Directory of Open Access Journals (Sweden)

    Reshu Saxena

    Full Text Available Nef is an accessory viral protein that promotes HIV-1 replication, facilitating alterations in cellular pathways via multiple protein-protein interactions. The advent of proteomics has expanded the focus on better identification of novel molecular pathways regulating disease progression. In this study, nef was sequenced from randomly selected patients, however, sequence variability identified did not elicited any specific mutation that could have segregated HIV-1 patients in different stages of disease progression. To explore the difference in Nef functionality based on sequence variability we used proteomics approach. Proteomic profiling was done to compare the effect of Nef variants in host cell protein expression. 2DGE in control and Nef transfected SupT1 cells demonstrated several differentially expressed proteins. Fourteen protein spots were detected with more than 1.5 fold difference. Significant down regulation was seen in six unique protein spots in the Nef treated cells. Proteins were identified as Cyclophilin A, EIF5A-1 isoform B, Rho GDI 1 isoform a, VDAC1, OTUB1 and α-enolase isoform 1 (ENO1 through LC-MS/MS. The differential expression of the 6 proteins was analyzed by Real time PCR, Western blotting and Immunofluorescence studies with two Nef variants (RP14 and RP01 in SupT1 cells. There was contrasting difference between the effect of these Nef variants upon the expression of these six proteins. Downregulation of α-enolase (ENO1, VDAC1 and OTUB1 was more significant by Nef RP01 whereas Cyclophilin A and RhoGDI were found to be more downregulated by Nef RP14. This difference in Nef variants upon host protein expression was also studied through a site directed mutant of Nef RP01 (55AAAAAAA61 and the effect was found to be reversed. Deciphering the role of these proteins mediated by Nef variants will open a new avenue of research in understanding Nef mediated pathogenesis. Overall study determines modulation of cellular protein

  5. Brain transcriptome-wide screen for HIV-1 Nef protein interaction partners reveals various membrane-associated proteins.

    Directory of Open Access Journals (Sweden)

    Ellen C Kammula

    Full Text Available HIV-1 Nef protein contributes essentially to the pathology of AIDS by a variety of protein-protein-interactions within the host cell. The versatile functionality of Nef is partially attributed to different conformational states and posttranslational modifications, such as myristoylation. Up to now, many interaction partners of Nef have been identified using classical yeast two-hybrid screens. Such screens rely on transcriptional activation of reporter genes in the nucleus to detect interactions. Thus, the identification of Nef interaction partners that are integral membrane proteins, membrane-associated proteins or other proteins that do not translocate into the nucleus is hampered. In the present study, a split-ubiquitin based yeast two-hybrid screen was used to identify novel membrane-localized interaction partners of Nef. More than 80% of the hereby identified interaction partners of Nef are transmembrane proteins. The identified hits are GPM6B, GPM6A, BAP31, TSPAN7, CYB5B, CD320/TCblR, VSIG4, PMEPA1, OCIAD1, ITGB1, CHN1, PH4, CLDN10, HSPA9, APR-3, PEBP1 and B3GNT, which are involved in diverse cellular processes like signaling, apoptosis, neurogenesis, cell adhesion and protein trafficking or quality control. For a subfraction of the hereby identified proteins we present data supporting their direct interaction with HIV-1 Nef. We discuss the results with respect to many phenotypes observed in HIV infected cells and patients. The identified Nef interaction partners may help to further elucidate the molecular basis of HIV-related diseases.

  6. Brain transcriptome-wide screen for HIV-1 Nef protein interaction partners reveals various membrane-associated proteins.

    Science.gov (United States)

    Kammula, Ellen C; Mötter, Jessica; Gorgels, Alexandra; Jonas, Esther; Hoffmann, Silke; Willbold, Dieter

    2012-01-01

    HIV-1 Nef protein contributes essentially to the pathology of AIDS by a variety of protein-protein-interactions within the host cell. The versatile functionality of Nef is partially attributed to different conformational states and posttranslational modifications, such as myristoylation. Up to now, many interaction partners of Nef have been identified using classical yeast two-hybrid screens. Such screens rely on transcriptional activation of reporter genes in the nucleus to detect interactions. Thus, the identification of Nef interaction partners that are integral membrane proteins, membrane-associated proteins or other proteins that do not translocate into the nucleus is hampered. In the present study, a split-ubiquitin based yeast two-hybrid screen was used to identify novel membrane-localized interaction partners of Nef. More than 80% of the hereby identified interaction partners of Nef are transmembrane proteins. The identified hits are GPM6B, GPM6A, BAP31, TSPAN7, CYB5B, CD320/TCblR, VSIG4, PMEPA1, OCIAD1, ITGB1, CHN1, PH4, CLDN10, HSPA9, APR-3, PEBP1 and B3GNT, which are involved in diverse cellular processes like signaling, apoptosis, neurogenesis, cell adhesion and protein trafficking or quality control. For a subfraction of the hereby identified proteins we present data supporting their direct interaction with HIV-1 Nef. We discuss the results with respect to many phenotypes observed in HIV infected cells and patients. The identified Nef interaction partners may help to further elucidate the molecular basis of HIV-related diseases.

  7. Determinants in HIV-1 Nef for enhancement of virus replication and depletion of CD4+ T lymphocytes in human lymphoid tissue ex vivo

    Directory of Open Access Journals (Sweden)

    Sertel Serkan

    2009-01-01

    Full Text Available Abstract Background HIV-1 Nef critically contributes to AIDS in part by augmenting virus titers in infected individuals. Analyzing which of Nef's activities contribute to HIV pathogenesis has been hampered by the lack of a cell culture model in which Nef exerts pronounced effects on HIV replication. The human lymphoid aggregate culture (HLAC from tonsil maintains the cell populations and cytokine milieu found in vivo, supports a productive infection without exogenous stimulation, and Nef contributes to efficient HIV-1 replication as well as CD4+ T cell depletion in this experimental ex vivo-model. Results To identify determinants in Nef that mediate these activities, we infected HLAC with a panel of isogenic HIV-1NL4-3 strains that encode for well-characterized mutants of HIV-1SF2 Nef. Determination of HIV-1 replication revealed that enhancement of the virus spread by Nef is governed by a complex set of protein interaction surfaces. In contrast, increased CD4+ T lymphocyte depletion depended on only two protein interaction surfaces in Nef that mediate either downregulation of cell surface CD4 or interaction with the NAKC signalosome. Consistently, in HLAC from 9 out of 14 donors, Nef enhanced CD4+ T cell depletion in the absence of a significant effect on virus replication. Moreover, our results suggest that this Nef-dependent enhancement in depletion occurred predominately in uninfected bystander CD4+ T cells. Conclusion Our findings suggest that Nef facilitates depletion of CD4+ T lymphocytes in HIV-1-infected lymphoid tissue ex vivo by increasing the pool of productively infected cells and by sensitizing bystander cells for killing. This ability might contribute to Nef's pathogenic potential in vivo.

  8. Cell-based Fluorescence Complementation Reveals a Role for HIV-1 Nef Protein Dimerization in AP-2 Adaptor Recruitment and CD4 Co-receptor Down-regulation.

    Science.gov (United States)

    Shu, Sherry T; Emert-Sedlak, Lori A; Smithgall, Thomas E

    2017-02-17

    The HIV-1 Nef accessory factor enhances viral infectivity, immune evasion, and AIDS progression. Nef triggers rapid down-regulation of CD4 via the endocytic adaptor protein 2 (AP-2) complex, a process linked to enhanced viral infectivity and immune escape. Here, we describe a bimolecular fluorescence complementation (BiFC) assay to visualize the interaction of Nef with AP-2 and CD4 in living cells. Interacting protein pairs were fused to complementary non-fluorescent fragments of YFP and co-expressed in 293T cells. Nef interactions with both CD4 and AP-2 resulted in complementation of YFP and a bright fluorescent signal by confocal microcopy that localized to the cell periphery. Co-expression of the AP-2 α subunit enhanced the Nef·AP-2 σ2 subunit BiFC signal and vice versa, suggesting that the AP-2 α-σ2 hemicomplex interacts cooperatively with Nef. Mutagenesis of Nef amino acids Arg-134, Glu-174, and Asp-175, which stabilize Nef for AP-2 α-σ2 binding in a recent co-crystal structure, substantially reduced AP-2 interaction without affecting CD4 binding. A dimerization-defective mutant of Nef failed to interact with either CD4 or AP-2 in the BiFC assay, indicating that Nef quaternary structure is required for CD4 and AP-2 recruitment as well as CD4 down-regulation. A small molecule previously shown to bind the Nef dimerization interface also reduced Nef interactions with AP-2 and CD4 and restored CD4 expression to the surface of HIV-infected cells. Our findings provide a mechanistic explanation for previous observations that dimerization-defective Nef mutants fail to down-regulate CD4 and validate the Nef dimerization interface as a target site for antiretroviral drug development. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  9. HIV-1 Nef binds the DOCK2-ELMO1 complex to activate rac and inhibit lymphocyte chemotaxis.

    Directory of Open Access Journals (Sweden)

    Ajit Janardhan

    2004-01-01

    Full Text Available The infectious cycle of primate lentiviruses is intimately linked to interactions between cells of the immune system. Nef, a potent virulence factor, alters cellular environments to increase lentiviral replication in the host, yet the mechanisms underlying these effects have remained elusive. Since Nef likely functions as an adaptor protein, we exploited a proteomic approach to directly identify molecules that Nef targets to subvert the signaling machinery in T cells. We purified to near homogeneity a major Nef-associated protein complex from T cells and identified by mass spectroscopy its subunits as DOCK2-ELMO1, a key activator of Rac in antigen- and chemokine-initiated signaling pathways, and Rac. We show that Nef activates Rac in T cell lines and in primary T cells following infection with HIV-1 in the absence of antigenic stimuli. Nef activates Rac by binding the DOCK2-ELMO1 complex, and this interaction is linked to the abilities of Nef to inhibit chemotaxis and promote T cell activation. Our data indicate that Nef targets a critical switch that regulates Rac GTPases downstream of chemokine- and antigen-initiated signaling pathways. This interaction enables Nef to influence multiple aspects of T cell function and thus provides an important mechanism by which Nef impacts pathogenesis by primate lentiviruses.

  10. Overlapping effector interfaces define the multiple functions of the HIV-1 Nef polyproline helix

    Directory of Open Access Journals (Sweden)

    Kuo Lillian S

    2012-05-01

    Full Text Available Abstract Background HIV-1 Nef is a multifunctional protein required for full pathogenicity of the virus. As Nef has no known enzymatic activity, it necessarily functions through protein-protein interaction interfaces. A critical Nef protein interaction interface is centered on its polyproline segment (P69VRPQVPLRP78 which contains the helical SH3 domain binding protein motif, PXXPXR. We hypothesized that any Nef-SH3 domain interactions would be lost upon mutation of the prolines or arginine of PXXPXR. Further, mutation of the non-motif “X” residues, (Q73, V74, and L75 would give altered patterns of inhibition for different Nef/SH3 domain protein interactions. Results We found that mutations of either of the prolines or the arginine of PXXPXR are defective for Nef-Hck binding, Nef/activated PAK2 complex formation and enhancement of virion infectivity (EVI. Mutation of the non-motif “X” residues (Q, V and L gave similar patterns of inhibition for Nef/activated PAK2 complex formation and EVI which were distinct from the pattern for Hck binding. These results implicate an SH3 domain containing protein other than Hck for Nef/activated PAK2 complex formation and EVI. We have also mutated Nef residues at the N-and C-terminal ends of the polyproline segment to explore interactions outside of PXXPXR. We discovered a new locus GFP/F (G67, F68, P69 and F90 that is required for Nef/activated PAK2 complex formation and EVI. MHC Class I (MHCI downregulation was only partially inhibited by mutating the PXXPXR motif residues, but was fully inhibited by mutating the C-terminal P78. Further, we observed that MHCI downregulation strictly requires G67 and F68. Our mutational analysis confirms the recently reported structure of the complex between Nef, AP-1 μ1 and the cytoplasmic tail of MHCI, but does not support involvement of an SH3 domain protein in MHCI downregulation. Conclusion Nef has evolved to be dependent on interactions with multiple SH3 domain

  11. Acute Viral Escape Selectively Impairs Nef-Mediated Major Histocompatibility Complex Class I Downmodulation and Increases Susceptibility to Antiviral T Cells.

    Science.gov (United States)

    Weiler, Andrea M; Das, Arpita; Akinyosoye, Oluwasayo; Cui, Sherry; O'Connor, Shelby L; Scheef, Elizabeth A; Reed, Jason S; Panganiban, Antonito T; Sacha, Jonah B; Rakasz, Eva G; Friedrich, Thomas C; Maness, Nicholas J

    2015-12-04

    Nef-specific CD8(+) T lymphocytes (CD8TL) are associated with control of simian immunodeficiency virus (SIV) despite extensive nef variation between and within animals. Deep viral sequencing of the immunodominant Mamu-B*017:01-restricted Nef165-173IW9 epitope revealed highly restricted evolution. A common acute escape variant, T170I, unexpectedly and uniquely degraded Nef's major histocompatibility complex class I (MHC-I) downregulatory capacity, rendering the virus more vulnerable to CD8TL targeting other epitopes. These data aid in a mechanistic understanding of Nef functions and suggest means of immunity-mediated control of lentivirus replication.

  12. Nef decreases HIV-1 sensitivity to neutralizing antibodies that target the membrane-proximal external region of TMgp41.

    Directory of Open Access Journals (Sweden)

    Rachel P J Lai

    2011-12-01

    Full Text Available Primate lentivirus nef is required for sustained virus replication in vivo and accelerated progression to AIDS. While exploring the mechanism by which Nef increases the infectivity of cell-free virions, we investigated a functional link between Nef and Env. Since we failed to detect an effect of Nef on the quantity of virion-associated Env, we searched for qualitative changes by examining whether Nef alters HIV-1 sensitivity to agents that target distinct features of Env. Nef conferred as much as 50-fold resistance to 2F5 and 4E10, two potent neutralizing monoclonal antibodies (nAbs that target the membrane proximal external region (MPER of TMgp41. In contrast, Nef had no effect on HIV-1 neutralization by MPER-specific nAb Z13e1, by the peptide inhibitor T20, nor by a panel of nAbs and other reagents targeting gp120. Resistance to neutralization by 2F5 and 4E10 was observed with Nef from a diverse range of HIV-1 and SIV isolates, as well as with HIV-1 virions bearing Env from CCR5- and CXCR4-tropic viruses, clade B and C viruses, or primary isolates. Functional analysis of a panel of Nef mutants revealed that this activity requires Nef myristoylation but that it is genetically separable from other Nef functions such as the ability to enhance virus infectivity and to downregulate CD4. Glycosylated-Gag from MoMLV substituted for Nef in conferring resistance to 2F5 and 4E10, indicating that this activity is conserved in a retrovirus that does not encode Nef. Given the reported membrane-dependence of MPER-recognition by 2F5 and 4E10, in contrast to the membrane-independence of Z13e1, the data here is consistent with a model in which Nef alters MPER recognition in the context of the virion membrane. Indeed, Nef and Glycosylated-Gag decreased the efficiency of virion capture by 2F5 and 4E10, but not by other nAbs. These studies demonstrate that Nef protects lentiviruses from one of the most broadly-acting classes of neutralizing antibodies. This newly

  13. HIV-1 Nef sequesters MHC-I intracellularly by targeting early stages of endocytosis and recycling.

    Science.gov (United States)

    Dirk, Brennan S; Pawlak, Emily N; Johnson, Aaron L; Van Nynatten, Logan R; Jacob, Rajesh A; Heit, Bryan; Dikeakos, Jimmy D

    2016-11-14

    A defining characteristic of HIV-1 infection is the ability of the virus to persist within the host. Specifically, MHC-I downregulation by the HIV-1 accessory protein Nef is of critical importance in preventing infected cells from cytotoxic T-cell mediated killing. Nef downregulates MHC-I by modulating the host membrane trafficking machinery, resulting in the endocytosis and eventual sequestration of MHC-I within the cell. In the current report, we utilized the intracellular protein-protein interaction reporter system, bimolecular fluorescence complementation (BiFC), in combination with super-resolution microscopy, to track the Nef/MHC-I interaction and determine its subcellular localization in cells. We demonstrate that this interaction occurs upon Nef binding the MHC-I cytoplasmic tail early during endocytosis in a Rab5-positive endosome. Disruption of early endosome regulation inhibited Nef-dependent MHC-I downregulation, demonstrating that Nef hijacks the early endosome to sequester MHC-I within the cell. Furthermore, super-resolution imaging identified that the Nef:MHC-I BiFC complex transits through both early and late endosomes before ultimately residing at the trans-Golgi network. Together we demonstrate the importance of the early stages of the endocytic network in the removal of MHC-I from the cell surface and its re-localization within the cell, which allows HIV-1 to optimally evade host immune responses.

  14. Down-regulation of CTLA-4 by HIV-1 Nef protein.

    Directory of Open Access Journals (Sweden)

    Mohamed El-Far

    Full Text Available HIV-1 Nef protein down-regulates several cell surface receptors through its interference with the cell sorting and trafficking machinery. Here we demonstrate for the first time the ability of Nef to down-regulate cell surface expression of the negative immune modulator CTLA-4. Down-regulation of CTLA-4 required the Nef motifs DD175, EE155 and LL165, all known to be involved in vesicle trafficking. Disruption of the lysosomal functions by pH-neutralizing agents prevented CTLA-4 down-regulation by Nef, demonstrating the implication of the endosomal/lysosomal compartments in this process. Confocal microscopy experiments visualized the co-localization between Nef and CTLA-4 in the early and recycling endosomes but not at the cell surface. Overall, our results provide a novel mechanism by which HIV-1 Nef interferes with the surface expression of the negative regulator of T cell activation CTLA-4. Down-regulation of CTLA-4 may contribute to the mechanisms by which HIV-1 sustains T cell activation, a critical step in viral replication and dissemination.

  15. HIV-1 Nef down-modulates C-C and C-X-C chemokine receptors via ubiquitin and ubiquitin-independent mechanism.

    Directory of Open Access Journals (Sweden)

    Prabha Chandrasekaran

    Full Text Available Human and Simian Immunodeficiency virus (HIV-1, HIV-2, and SIV encode an accessory protein, Nef, which is a pathogenesis and virulence factor. Nef is a multivalent adapter that dysregulates the trafficking of many immune cell receptors, including chemokine receptors (CKRs. Physiological endocytic itinerary of agonist occupied CXCR4 involves ubiquitinylation of the phosphorylated receptor at three critical lysine residues and dynamin-dependent trafficking through the ESCRT pathway into lysosomes for degradation. Likewise, Nef induced CXCR4 degradation was critically dependent on the three lysines in the C-terminal -SSLKILSKGK- motif. Nef directly recruits the HECT domain E3 ligases AIP4 or NEDD4 to CXCR4 in the resting state. This mechanism was confirmed by ternary interactions of Nef, CXCR4 and AIP4 or NEDD4; by reversal of Nef effect by expression of catalytically inactive AIP4-C830A mutant; and siRNA knockdown of AIP4, NEDD4 or some ESCRT-0 adapters. However, ubiquitinylation dependent lysosomal degradation was not the only mechanism by which Nef downregulated CKRs. Agonist and Nef mediated CXCR2 (and CXCR1 degradation was ubiquitinylation independent. Nef also profoundly downregulated the naturally truncated CXCR4 associated with WHIM syndrome and engineered variants of CXCR4 that resist CXCL12 induced internalization via an ubiquitinylation independent mechanism.

  16. Species-specific activity of SIV Nef and HIV-1 Vpu in overcoming restriction by tetherin/BST2.

    Directory of Open Access Journals (Sweden)

    Bin Jia

    2009-05-01

    Full Text Available Tetherin, also known as BST2, CD317 or HM1.24, was recently identified as an interferon-inducible host-cell factor that interferes with the detachment of virus particles from infected cells. HIV-1 overcomes this restriction by expressing an accessory protein, Vpu, which counteracts tetherin. Since lentiviruses of the SIV(smm/mac/HIV-2 lineage do not have a vpu gene, this activity has likely been assumed by other viral gene products. We found that deletion of the SIV(mac239 nef gene significantly impaired virus release in cells expressing rhesus macaque tetherin. Virus release could be restored by expressing Nef in trans. However, Nef was unable to facilitate virus release in the presence of human tetherin. Conversely, Vpu enhanced virus release in the presence of human tetherin, but not in the presence of rhesus tetherin. In accordance with the species-specificity of Nef in mediating virus release, SIV Nef downregulated cell-surface expression of rhesus tetherin, but did not downregulate human tetherin. The specificity of SIV Nef for rhesus tetherin mapped to four amino acids in the cytoplasmic domain of the molecule that are missing from human tetherin, whereas the specificity of Vpu for human tetherin mapped to amino acid differences in the transmembrane domain. Nef alleles of SIV(smm, HIV-2 and HIV-1 were also able to rescue virus release in the presence of both rhesus macaque and sooty mangabey tetherin, but were generally ineffective against human tetherin. Thus, the ability of Nef to antagonize tetherin from these Old World primates appears to be conserved among the primate lentiviruses. These results identify Nef as the viral gene product of SIV that opposes restriction by tetherin in rhesus macaques and sooty mangabeys, and reveal species-specificity in the activities of both Nef and Vpu in overcoming tetherin in their respective hosts.

  17. The HIV-1 Nef protein and phagocyte NADPH oxidase activation

    DEFF Research Database (Denmark)

    Vilhardt, Frederik; Plastre, Olivier; Sawada, Makoto;

    2002-01-01

    -regulation of phagocyte NADPH oxidase subunits. Nef mutants lacking motifs involved in the interaction with Vav and PAK failed to reproduce the effects of wild type Nef, suggesting a role for the Vav/Rac/PAK signaling pathway. The following results suggest a key role for Rac in the priming effect of Nef. (i) Inactivation...... of Rac by Clostridium difficile toxin B abolished the Nef effect. (ii) The fraction of activated Rac1 was increased in Nef-transduced cells, and (iii) the dominant positive Rac1(V12) mutant mimicked the effect of Nef. These results are to our knowledge the first analysis of the effect of Rac activation...

  18. Self-association of the Lentivirus protein, Nef

    Directory of Open Access Journals (Sweden)

    Garcia J Victor

    2010-09-01

    Full Text Available Abstract Background The HIV-1 pathogenic factor, Nef, is a multifunctional protein present in the cytosol and on membranes of infected cells. It has been proposed that a spatial and temporal regulation of the conformation of Nef sequentially matches Nef's multiple functions to the process of virion production. Further, it has been suggested that dimerization is required for multiple Nef activities. A dimerization interface has been proposed based on intermolecular contacts between Nefs within hexagonal Nef/FynSH3 crystals. The proposed dimerization interface consists of the hydrophobic B-helix and flanking salt bridges between R105 and D123. Here, we test whether Nef self-association is mediated by this interface and address the overall significance of oligomerization. Results By co-immunoprecipitation assays, we demonstrated that HIV-1Nef exists as monomers and oligomers with about half of the Nef protomers oligomerized. Nef oligomers were found to be present in the cytosol and on membranes. Removal of the myristate did not enhance the oligomerization of soluble Nef. Also, SIVNef oligomerizes despite lacking a dimerization interface functionally homologous to that proposed for HIV-1Nef. Moreover, HIV-1Nef and SIVNef form hetero-oligomers demonstrating the existence of homologous oligomerization interfaces that are distinct from that previously proposed (R105-D123. Intracellular cross-linking by formaldehyde confirmed that SF2Nef dimers are present in intact cells, but surprisingly self-association was dependent on R105, but not D123. SIVMAC239Nef can be cross-linked at its only cysteine, C55, and SF2Nef is also cross-linked, but at C206 instead of C55, suggesting that Nefs exhibit multiple dimeric structures. ClusPro dimerization analysis of HIV-1Nef homodimers and HIV-1Nef/SIVNef heterodimers identified a new potential dimerization interface, including a dibasic motif at R105-R106 and a six amino acid hydrophobic surface. Conclusions We have

  19. One hundred years and still counting: the story of NEF--yesterday, today, and tomorrow.

    Science.gov (United States)

    Bowar-Ferres, Susan L; Fitzpatrick, M Louise; McClure, Margaret L

    2014-01-01

    Nurses Educational Funds, Inc (NEF) is the oldest and largest professionally endorsed source of funds for advanced study in nursing, which celebrated its Centennial in 2012. This philanthropic nonprofit organization is notable for its roots; NEF was organized by nurses specifically for nurses. Its history dates back to 1912, when it began in memory of Isabel Hampton Robb at Teacher's College, where the first graduate nursing education programs began. The initial Robb Memorial Fund was incorporated as a nonprofit organization in 1941 and officially became Nursing Educational Funds, Inc, in 1954. The NEF's sole mission is to raise money and give it for graduate-level scholarships in nursing education, service, practice, and research. Since its origin, more than 1000 doctoral and master's students from a broad array of schools across the nation have been recipients of awards. The NEF Board is a totally volunteer, highly dedicated group of nursing, business, and other professional leaders, who are steadfastly committed to this critical effort. Scholarships for graduate nursing education are imperative to meet the need to grow the pipeline of faculty. As charged by the 2010 Institute of Medicine report, the goal to increase the number of baccalaureate nurses to 80% of the workforce and to double the number of nurses with doctoral degrees both by 2020 speak to the heart of NEF. Funds raised currently are largely from Board members, individual donors, modest foundational support, and a number of bequests. As the nursing population grows older, the potential for bequests or planned giving becomes a realistic goal. Former NEF scholars have not unfortunately been a financial source, although pay back is an expectation. Nurses are the best ones to tell this compelling story to corporations and foundations as NEF continues to persist in the commitment to support graduate nursing education.

  20. A Conserved GPG-Motif in the HIV-1 Nef Core Is Required for Principal Nef-Activities.

    Directory of Open Access Journals (Sweden)

    Marta Martínez-Bonet

    Full Text Available To find out new determinants required for Nef activity we performed a functional alanine scanning analysis along a discrete but highly conserved region at the core of HIV-1 Nef. We identified the GPG-motif, located at the 121-137 region of HIV-1 NL4.3 Nef, as a novel protein signature strictly required for the p56Lck dependent Nef-induced CD4-downregulation in T-cells. Since the Nef-GPG motif was dispensable for CD4-downregulation in HeLa-CD4 cells, Nef/AP-1 interaction and Nef-dependent effects on Tf-R trafficking, the observed effects on CD4 downregulation cannot be attributed to structure constraints or to alterations on general protein trafficking. Besides, we found that the GPG-motif was also required for Nef-dependent inhibition of ring actin re-organization upon TCR triggering and MHCI downregulation, suggesting that the GPG-motif could actively cooperate with the Nef PxxP motif for these HIV-1 Nef-related effects. Finally, we observed that the Nef-GPG motif was required for optimal infectivity of those viruses produced in T-cells. According to these findings, we propose the conserved GPG-motif in HIV-1 Nef as functional region required for HIV-1 infectivity and therefore with a potential interest for the interference of Nef activity during HIV-1 infection.

  1. Lack of differences in HIV-1 nef functional domains in infected chinese blood donors at different stages of disease.

    Science.gov (United States)

    Chen, Yue; Caruso, Lori; Shen, Chengli; Wu, Hao; Zhou, Yushen; Gupta, Phalguni

    2007-09-01

    HIV-1 nef regions were amplified by polymerase chain reaction and sequenced from DNA samples of five asymptomatic subjects and five AIDS patients from a cohort of HIV-1-infected Chinese plasma and blood donors. Sequence analysis revealed that regardless of the stage of disease, each patient's HIV-1 nef sequences belonged to the clade B' subtype. Although there are some differences between the sequences from different patients, no significant differences have been detected in nef nucleotide sequences or functional motifs in the deduced amino acid sequences from patients at different stages of the disease. Furthermore, the predicted binding motifs of HLA-A2 and HLA-A11 were highly conserved among patient nef sequences. These results will contribute to a better understanding of the pathogenesis of circulating HIV-1 in infected Chinese former blood donors and may have important implications in developing an epitope-based vaccine suitable for Chinese blood donors.

  2. Mutation of a diacidic motif in SIV-PBj Nef impairs T-cell activation and enteropathic disease

    Directory of Open Access Journals (Sweden)

    Prüfer Steffen

    2011-03-01

    Full Text Available Abstract Background The non-pathogenic course of SIV infection in its natural host is characterized by robust viral replication in the absence of chronic immune activation and T cell proliferation. In contrast, acutely lethal enteropathic SIVsmm strain PBj induces a strong immune activation and causes a severe acute and lethal disease in pig-tailed macaques after cross-species transmission. One important pathogenicity factor of the PBj virus is the PBj-Nef protein, which contains a conserved diacidic motif and, unusually, an immunoreceptor tyrosine-based activation motif (ITAM. Results Mutation of the diacidic motif in the Nef protein of the SIVsmmPBj abolishes the acute phenotype of this virus. In vitro, wild-type and mutant PBj (PBj-Nef202/203GG viruses replicated to similar levels in macaque PBMCs, but PBj-Nef202/203GG no longer triggers ERK mitogen-activated protein (MAP kinase pathway including an alteration of a Nef-associated Raf-1/ERK-2 multiprotein signaling complex. Moreover, stimulation of IL-2 and down-modulation of CD4 and CD28 were impaired in the mutant virus. Pig-tailed macaques infected with PBj-Nef202/203GG did not show enteropathic complications and lethality as observed with wild-type PBj virus, despite efficient replication of both viruses in vivo. Furthermore, PBj-Nef202/203GG infected animals revealed reduced T-cell activation in periphery lymphoid organs and no detectable induction of IL-2 and IL-6. Conclusions In sum, we report here that mutation of the diacidic motif in the PBj-Nef protein abolishes disease progression in pig-tailed macaques despite efficient replication. These data suggest that alterations in the ability of a lentivirus to promote T cell activation and proliferation can have a dramatic impact on its pathogenic potential.

  3. Identification of a highly conserved valine-glycine-phenylalanine amino acid triplet required for HIV-1 Nef function

    Directory of Open Access Journals (Sweden)

    Meuwissen Pieter J

    2012-04-01

    Full Text Available Abstract Background The Nef protein of HIV facilitates virus replication and disease progression in infected patients. This role as pathogenesis factor depends on several genetically separable Nef functions that are mediated by interactions of highly conserved protein-protein interaction motifs with different host cell proteins. By studying the functionality of a series of nef alleles from clinical isolates, we identified a dysfunctional HIV group O Nef in which a highly conserved valine-glycine-phenylalanine (VGF region, which links a preceding acidic cluster with the following proline-rich motif into an amphipathic surface was deleted. In this study, we aimed to study the functional importance of this VGF region. Results The dysfunctional HIV group O8 nef allele was restored to the consensus sequence, and mutants of canonical (NL4.3, NA-7, SF2 and non-canonical (B2 and C1422 HIV-1 group M nef alleles were generated in which the amino acids of the VGF region were changed into alanines (VGF→AAA and tested for their capacity to interfere with surface receptor trafficking, signal transduction and enhancement of viral replication and infectivity. We found the VGF motif, and each individual amino acid of this motif, to be critical for downregulation of MHC-I and CXCR4. Moreover, Nef’s association with the cellular p21-activated kinase 2 (PAK2, the resulting deregulation of cofilin and inhibition of host cell actin remodeling, and targeting of Lck kinase to the trans-golgi-network (TGN were affected as well. Of particular interest, VGF integrity was essential for Nef-mediated enhancement of HIV virion infectivity and HIV replication in peripheral blood lymphocytes. For targeting of Lck kinase to the TGN and viral infectivity, especially the phenylalanine of the triplet was essential. At the molecular level, the VGF motif was required for the physical interaction of the adjacent proline-rich motif with Hck. Conclusion Based on these findings, we

  4. HIV-1 Nef is transferred from expressing T cells to hepatocytic cells through conduits and enhances HCV replication.

    Directory of Open Access Journals (Sweden)

    In-Woo Park

    Full Text Available HIV-1 infection enhances HCV replication and as a consequence accelerates HCV-mediated hepatocellular carcinoma (HCC. However, the precise molecular mechanism by which this takes place is currently unknown. Our data showed that infectious HIV-1 failed to replicate in human hepatocytic cell lines. No discernible virus replication was observed, even when the cell lines transfected with HIV-1 proviral DNA were co-cultured with Jurkat T cells, indicating that the problem of liver deterioration in the co-infected patient is not due to the replication of HIV-1 in the hepatocytes of the HCV infected host. Instead, HIV-1 Nef protein was transferred from nef-expressing T cells to hepatocytic cells through conduits, wherein up to 16% (average 10% of the cells harbored the transferred Nef, when the hepatocytic cells were co-cultured with nef-expressing Jurkat cells for 24 h. Further, Nef altered the size and numbers of lipid droplets (LD, and consistently up-regulated HCV replication by 1.5∼2.5 fold in the target subgenomic replicon cells, which is remarkable in relation to the initially indolent viral replication. Nef also dramatically augmented reactive oxygen species (ROS production and enhanced ethanol-mediated up-regulation of HCV replication so as to accelerate HCC. Taken together, these data indicate that HIV-1 Nef is a critical element in accelerating progression of liver pathogenesis via enhancing HCV replication and coordinating modulation of key intra- and extra-cellular molecules for liver decay.

  5. An MHC-I cytoplasmic domain/HIV-1 Nef fusion protein binds directly to the mu subunit of the AP-1 endosomal coat complex.

    Directory of Open Access Journals (Sweden)

    Rajendra Kumar Singh

    Full Text Available BACKGROUND: The down-regulation of the major histocompatibility complex class I (MHC-I from the surface of infected cells by the Nef proteins of primate immunodeficiency viruses likely contributes to pathogenesis by providing evasion of cell-mediated immunity. HIV-1 Nef-induced down-regulation involves endosomal trafficking and a cooperative interaction between the cytoplasmic domain (CD of MHC-I, Nef, and the clathrin adaptor protein complex-1 (AP-1. The CD of MHC-I contains a key tyrosine within the sequence YSQA that is required for down-regulation by Nef, but this sequence does not conform to the canonical AP-binding tyrosine-based motif Yxxphi, which mediates binding to the medium (micro subunits of AP complexes. We previously proposed that Nef allows the MHC-I CD to bind the mu subunit of AP-1 (micro1 as if it contained a Yxxphimotif. METHODS AND FINDINGS: Here, we show that a direct interaction between the MHC-I CD/Nef and micro1 plays a primary role in the down-regulation of MHC-I: GST pulldown assays using recombinant proteins indicated that most of the MHC-I CD and Nef residues that are required for the down-regulation in human cells contribute to direct interactions with a truncated version of micro1. Specifically, the tyrosine residue of the YSQA sequence in the MHC-I CD as well as Nef residues E62-65 and P78 each contributed to the interaction between MHC-I CD/Nef and micro1 in vitro, whereas Nef M20 had little to no role. Conversely, residues F172/D174 and V392/L395 of the binding pocket on micro1 for Yxxphi motifs were required for a robust interaction. CONCLUSIONS: These data indicate that the MHC-I cytoplasmic domain, Nef, and the C-terminal two thirds of the mu subunit of AP-1 are sufficient to constitute a biologically relevant interaction. The data also reveal an unexpected role for a hydrophobic pocket in micro1 for interaction with MHC-I CD/Nef.

  6. Attenuation of multiple Nef functions in HIV-1 elite controllers

    Directory of Open Access Journals (Sweden)

    Mwimanzi Philip

    2013-01-01

    Full Text Available Abstract Background Impaired HIV-1 Gag, Pol, and Env function has been described in elite controllers (EC who spontaneously suppress plasma viremia to Results In general, EC Nef clones were functional; however, all five activities were significantly lower in EC compared to CP. Nef clones from HLA-B*57-expressing EC exhibited poorer CD4 down-regulation function compared to those from non-B*57 EC, and the number of EC-specific B*57-associated Nef polymorphisms correlated inversely with 4 of 5 Nef functions in these individuals. Conclusion Results indicate that decreased HIV-1 Nef function, due in part to host immune selection pressures, may be a hallmark of the EC phenotype.

  7. [Linking Nef with the immunological and virological determinants of AIDS].

    Science.gov (United States)

    Gómez-Icazbalceta, Guillermo; Larralde, Carlos

    2009-01-01

    Nef is an accessory protein from HIV-1 involved in viral replication, depletion of CD4 + T cells and progression to AIDS. The participation of Nef in altering several of the cellular and subcellular components has been studied in detail. The finding that infected patients over 15 years with HIV-1 variants that expressed a defective nef gene had not progressed to AIDS, suggested that Nef played a decisive role in the outcome of AIDS, through the regulation of different virological and immunological components. In addition, the mere expression of Nef is able to induce AIDS in transgenic mice and alter the kinetics of replication of the virus in infected monkeys. This review examines the relationship between the expression of Nef and the ultimate consequences of HIV-1 infection, taking into account the most relevant clinical parameters: viral load, counting of CD4 + T cells and progression to AIDS. Under this approach, it is emphasized the role of Nef as immunomodulator and also as independent mediator of damage to the immune system.

  8. HIV-1 Nef breaches placental barrier in rat model.

    Science.gov (United States)

    Singh, Poonam; Agnihotri, Saurabh Kumar; Tewari, Mahesh Chandra; Kumar, Sadan; Sachdev, Monika; Tripathi, Raj Kamal

    2012-01-01

    The vertical transmission of HIV-1 from the mother to fetus is known, but the molecular mechanism regulating this transmission is not fully characterized. The fetus is highly protected by the placenta, which does not permit microbial pathogens to cross the placental barrier. In the present study, a rat model was established to observe the effect of HIV-1 protein Nef on placental barrier. Evans blue dye was used to assay permeability of placental barrier and fourteen day pregnant Sprague Dawley rats were injected intravenously with 2% Evans blue dye along with various concentrations of recombinant Nef. After an hour, animals were sacrificed and dye migration was observed through the assimilation of peripheral blood into fetus. Interestingly, traces of recombinant Nef protein were detected in the embryo as well as amniotic fluid and amniotic membrane along with placenta and uterus. Our study indicates that recombinant HIV-1-Nef protein breaches the placental barrier and allows the migration of Evans blue dye to the growing fetus. Further the concentration of Nef protein in blood is directly proportional to the intensity of dye migration and to the amount of Nef protein detected in uterus, placenta, amniotic membrane, amniotic fluid and embryo. Based on this study, it can be concluded that the HIV-1 Nef protein has a direct effect on breaching of the placental barrier in the model we have established in this study. Our observations will be helpful to understand the molecular mechanisms related to this breach of placental barrier by Nef in humans and may be helpful to identify specific Nef inhibitors.

  9. Immunogenic compositions comprising human immunodeficiency virus (HIV) mosaic Nef proteins

    Science.gov (United States)

    Korber, Bette T [Los Alamos, NM; Perkins, Simon [Los Alamos, NM; Bhattacharya, Tanmoy [Los Alamos, NM; Fischer, William M [Los Alamos, NM; Theiler, James [Los Alamos, NM; Letvin, Norman [Boston, MA; Haynes, Barton F [Durham, NC; Hahn, Beatrice H [Birmingham, AL; Yusim, Karina [Los Alamos, NM; Kuiken, Carla [Los Alamos, NM

    2012-02-21

    The present invention relates to mosaic clade M HIV-1 Nef polypeptides and to compositions comprising same. The polypeptides of the invention are suitable for use in inducing an immune response to HIV-1 in a human.

  10. On projective space bundle with nef normalized tautological line bundle

    CERN Document Server

    Yasutake, Kazunori

    2011-01-01

    In this paper, we study the structure of projective space bundles whose relative anti-canonical line bundle is nef. As an application, we get a characterization of abelian varieties up to finite etale covering.

  11. A Truncated Nef Peptide from SIVcpz Inhibits the Production of HIV-1 Infectious Progeny

    Directory of Open Access Journals (Sweden)

    Marcela Sabino Cunha

    2016-07-01

    Full Text Available Nef proteins from all primate Lentiviruses, including the simian immunodeficiency virus of chimpanzees (SIVcpz, increase viral progeny infectivity. However, the function of Nef involved with the increase in viral infectivity is still not completely understood. Nonetheless, until now, studies investigating the functions of Nef from SIVcpz have been conducted in the context of the HIV-1 proviruses. In an attempt to investigate the role played by Nef during the replication cycle of an SIVcpz, a Nef-defective derivative was obtained from the SIVcpzWTGab2 clone by introducing a frame shift mutation at a unique restriction site within the nef sequence. This nef-deleted clone expresses an N-terminal 74-amino acid truncated peptide of Nef and was named SIVcpz-tNef. We found that the SIVcpz-tNef does not behave as a classic nef-deleted HIV-1 or simian immunodeficiency virus of macaques SIVmac. Markedly, SIVcpz-tNef progeny from both Hek-293T and Molt producer cells were completely non-infectious. Moreover, the loss in infectivity of SIVcpz-tNef correlated with the inhibition of Gag and GagPol processing. A marked accumulation of Gag and very low levels of reverse transcriptase were detected in viral lysates. Furthermore, these observations were reproduced once the tNef peptide was expressed in trans both in SIVcpzΔNef and HIV-1WT expressing cells, demonstrating that the truncated peptide is a dominant negative for viral processing and infectivity for both SIVcpz and HIV-1. We demonstrated that the truncated Nef peptide binds to GagPol outside the protease region and by doing so probably blocks processing of both GagPol and Gag precursors at a very early stage. This study demonstrates for the first time that naturally-occurring Nef peptides can potently block lentiviral processing and infectivity.

  12. A Truncated Nef Peptide from SIVcpz Inhibits the Production of HIV-1 Infectious Progeny

    Science.gov (United States)

    Sabino Cunha, Marcela; Lima Sampaio, Thatiane; Peterlin, B. Matija; Jesus da Costa, Luciana

    2016-01-01

    Nef proteins from all primate Lentiviruses, including the simian immunodeficiency virus of chimpanzees (SIVcpz), increase viral progeny infectivity. However, the function of Nef involved with the increase in viral infectivity is still not completely understood. Nonetheless, until now, studies investigating the functions of Nef from SIVcpz have been conducted in the context of the HIV-1 proviruses. In an attempt to investigate the role played by Nef during the replication cycle of an SIVcpz, a Nef-defective derivative was obtained from the SIVcpzWTGab2 clone by introducing a frame shift mutation at a unique restriction site within the nef sequence. This nef-deleted clone expresses an N-terminal 74-amino acid truncated peptide of Nef and was named SIVcpz-tNef. We found that the SIVcpz-tNef does not behave as a classic nef-deleted HIV-1 or simian immunodeficiency virus of macaques SIVmac. Markedly, SIVcpz-tNef progeny from both Hek-293T and Molt producer cells were completely non-infectious. Moreover, the loss in infectivity of SIVcpz-tNef correlated with the inhibition of Gag and GagPol processing. A marked accumulation of Gag and very low levels of reverse transcriptase were detected in viral lysates. Furthermore, these observations were reproduced once the tNef peptide was expressed in trans both in SIVcpzΔNef and HIV-1WT expressing cells, demonstrating that the truncated peptide is a dominant negative for viral processing and infectivity for both SIVcpz and HIV-1. We demonstrated that the truncated Nef peptide binds to GagPol outside the protease region and by doing so probably blocks processing of both GagPol and Gag precursors at a very early stage. This study demonstrates for the first time that naturally-occurring Nef peptides can potently block lentiviral processing and infectivity. PMID:27399760

  13. Nef functions in BLT mice to enhance HIV-1 replication and deplete CD4+CD8+ thymocytes

    Directory of Open Access Journals (Sweden)

    Zou Wei

    2012-05-01

    Full Text Available Abstract Background The outcome of untreated HIV-1 infection is progression to AIDS and death in nearly all cases. Some important exceptions are the small number of patients infected with HIV-1 deleted for the accessory gene, nef. With these infections, disease progression is entirely suppressed or greatly delayed. Whether Nef is critical for high levels of replication or is directly cytotoxic remains controversial. The major problem in determining the role of Nef in HIV/AIDS has been the lack of tractable in vivo models where Nef’s complex pathogenic phenotype can be recapitulated. Results Intravenous inoculation (3000 to 600,000 TCIU of BLT humanized mice with HIV-1LAI reproducibly establishes a systemic infection. HIV-1LAI (LAI replicates to high levels (peak viral load in blood 8,200,000 ± 1,800,000 copies of viral RNA/ml, range 3,600,000 to 20,400,000; n = 9 and exhaustively depletes CD4+ T cells in blood and tissues. CD4+CD8+ thymocytes were also efficiently depleted but CD4+CD8- thymocytes were partially resistant to cell killing by LAI. Infection with a nef-deleted LAI (LAINefdd gave lower peak viral loads (1,220,000 ± 330,000, range 27,000 to 4,240,000; n = 17. For fourteen of seventeen LAINefdd-infected mice, there was little to no loss of either CD4+ T cells or thymocytes. Both LAI- and LAINefdd-infected mice had about 8% of total peripheral blood CD8+ T cells that were CD38+HLA-DR+ compared dd-infected mice that lost CD4+ T cells received 600,000 TCIU. All three exhibited peak viral loads over 3,000,000 copies of LAINefdd RNA/ml. Over an extended time course, substantial systemic CD4+ T cell loss was observed for the three mice, but there was no loss of CD4+CD8+ or CD4+CD8- thymocytes. Conclusion We conclude Nef is necessary for elevated viral replication and as a result indirectly contributes to CD4+ T cell killing. Further, Nef was not necessary for the activation of peripheral blood CD8+ T cells following

  14. T cell activation and proliferation characteristic for HIV-Nef transgenic mice is lymphopenia induced

    NARCIS (Netherlands)

    Koenen, Paul G. F.; Hofhuis, Frans M.; Osterwegel, Mariette A.; Tesselaar, Kiki

    2007-01-01

    The HIV-Nef protein has been implicated in generating high viral loads and T cell activation. Transgenic (tg) mice with constitutive T cell-specific Nef expression show a dramatic reduction in T cell number and highly increased T cell turnover. Previous studies in Nef tg mice attributed this T cell

  15. Nefness of adjoint bundles for ample vector bundles

    Directory of Open Access Journals (Sweden)

    Hidetoshi Maeda

    1995-11-01

    Full Text Available Let E be an ample vector bundle of rank >1 on a smooth complex projective variety X of dimension n. This paper gives a classification of pairs (X,E whose adjoint bundles K_X+det E are not nef in the case when  r=n-2.

  16. Human Leukocyte Antigen (HLA Class I Down-Regulation by Human Immunodeficiency Virus Type 1 Negative Factor (HIV-1 Nef: What Might We Learn From Natural Sequence Variants?

    Directory of Open Access Journals (Sweden)

    Philip Mwimanzi

    2012-09-01

    Full Text Available HIV-1 causes a chronic infection in humans that is characterized by high plasma viremia, progressive loss of CD4+ T lymphocytes, and severe immunodeficiency resulting in opportunistic disease and AIDS. Viral persistence is mediated in part by the ability of the Nef protein to down-regulate HLA molecules on the infected cell surface, thereby allowing HIV-1 to evade recognition by antiviral CD8+ T lymphocytes. Extensive research has been conducted on Nef to determine protein domains that are required for its immune evasion activities and to identify critical cellular co-factors, and our mechanistic understanding of this process is becoming more complete. This review highlights our current knowledge of Nef-mediated HLA class I down-regulation and places this work in the context of naturally occurring sequence variation in this protein. We argue that efforts to fully understand the critical role of Nef for HIV-1 pathogenesis will require greater analysis of patient-derived sequences to elucidate subtle differences in immune evasion activity that may alter clinical outcome.

  17. Astrocytic expression of HIV-1 Nef impairs spatial and recognition memory

    OpenAIRE

    2012-01-01

    Despite the widespread use of antiretroviral therapy that effectively limits viral replication, memory impairment remains a dilemma for HIV infected people. In the CNS, HIV infection of astrocytes leads to the production of the HIV-1 Nef protein without viral replication. Post mortem studies have found Nef expression in hippocampal astrocytes of people with HIV associated dementia suggesting that astrocytic Nef may contribute to HIV associated cognitive impairment even when viral replication ...

  18. Natural variation of the nef gene in human immunodeficiency virus type 2 infections in Portugal

    OpenAIRE

    Pádua, E.; Jenkins, A.; S. Brown; Bootman, J.; Paixão, M T; Almond, N; Berry, N

    2003-01-01

    Human immunodeficiency virus type 2 (HIV-2) infections cause severe immunodeficiency in humans, although HIV-2 is associated frequently with reduced virulence and pathogenicity compared to HIV-1. Genetic determinants that play a role in HIV pathogenesis are relatively poorly understood but nef has been implicated in inducing a more pathogenic phenotype in vivo. However, relatively little is known about the role of nef in HIV-2 pathogenesis. To address this, the genetic composition of 44 nef a...

  19. Link between Primate Lentiviral Coreceptor Usage and Nef Function

    Directory of Open Access Journals (Sweden)

    Jan Schmökel

    2013-11-01

    Full Text Available Simian immunodeficiency virus (SIVsmm infection of sooty mangabeys (Cercocebus atys is characterized by stable CD4+ T cell counts despite high plasma levels of CCR5-tropic viruses. However, in rare instances, SIVsmm acquires CXCR4 coreceptor tropism and causes severe CD4+ T cell depletion, albeit without clinical signs of immunodeficiency. Here, we show that CXCR4-tropic SIVsmm strains lost their ability to downmodulate TCR-CD3 by evolving unusual Nef mutations that initially reduced (I132V and subsequently disrupted (I123L and L146F interaction with the CD3 ζ chain. This coevolution of Env and Nef function suggests that CD3 downmodulation is advantageous for viral replication in activated CCR5+ memory T cells, but not in resting naive CXCR4+ T cells that have not yet undergone TCR-CD3-mediated stimulation. This may explain why HIV-1, which generally lacks the CD3 downmodulation function, commonly switches to CXCR4 usage, whereas this is extremely rare for SIV strains that have retained this Nef activity.

  20. Astrocytic expression of HIV-1 Nef impairs spatial and recognition memory.

    Science.gov (United States)

    Chompre, Gladys; Cruz, Emmanuel; Maldonado, Lucianette; Rivera-Amill, Vanessa; Porter, James T; Noel, Richard J

    2013-01-01

    Despite the widespread use of antiretroviral therapy that effectively limits viral replication, memory impairment remains a dilemma for HIV infected people. In the CNS, HIV infection of astrocytes leads to the production of the HIV-1 Nef protein without viral replication. Post mortem studies have found Nef expression in hippocampal astrocytes of people with HIV associated dementia suggesting that astrocytic Nef may contribute to HIV associated cognitive impairment even when viral replication is suppressed. To test whether astrocytic expression of Nef is sufficient to induce cognitive deficits, we examined the effect of implanting primary rat astrocytes expressing Nef into the hippocampus on spatial and recognition memory. Rats implanted unilaterally with astrocytes expressing Nef showed impaired novel location and novel object recognition in comparison with controls implanted with astrocytes expressing green fluorescent protein (GFP). This impairment was correlated with an increase in chemokine ligand 2 (CCL2) expression and the infiltration of peripheral macrophages into the hippocampus at the site of injection. Furthermore, the Nef exposed rats exhibited a bilateral loss of CA3 neurons. These results suggest that Nef protein expressed by the implanted astrocytes activates the immune system leading to neuronal damage and spatial and recognition memory deficits. Therefore, the continued expression of Nef by astrocytes in the absence of viral replication has the potential to contribute to HIV associated cognitive impairment.

  1. Cysteine 138 mutation in HIV-1 Nef from patients with delayed disease progression

    DEFF Research Database (Denmark)

    Tolstrup, Martin; Laursen, Alex Lund; Gerstoft, J.;

    2006-01-01

    on the delayed disease status. However, the results demonstrate a high incidence of a single amino acid polymorphism (cysteine 138) in HIV-1 Nef. The allelic frequency of cysteine 138 between the delayed disease progression group and the progressor group was found to be statistically significant (P = 0.......0139). The phylogeny of isolates was investigated and the variants harbouring the cysteine 138 mutation clustered independently. CONCLUSION: The present study describes a viral genetic polymorphism related to AIDS disease progression. The polymorphism (cysteine 138) has previously been reported to confer decreased...... viral replication (Premkumar DR, et al. AIDS Res Hum Retroviruses 1996; 12(4): 337-45). A sequence database search for comparative mutations revealed a high frequency of cysteine 138 in patients with reported SP AIDS...

  2. Construction of Nef-positive doxycycline-dependent HIV-1 variants using bicistronic expression elements

    Energy Technology Data Exchange (ETDEWEB)

    Velden, Yme U. van der; Kleibeuker, Wendy; Harwig, Alex; Klaver, Bep; Siteur-van Rijnstra, Esther; Frankin, Esmay; Berkhout, Ben; Das, Atze T., E-mail: a.t.das@amc.uva.nl

    2016-01-15

    Conditionally replicating HIV-1 variants that can be switched on and off at will are attractive tools for HIV research. We previously developed a genetically modified HIV-1 variant that replicates exclusively when doxycycline (dox) is administered. The nef gene in this HIV-rtTA variant was replaced with the gene encoding the dox-dependent rtTA transcriptional activator. Because loss of Nef expression compromises virus replication in primary cells and precludes studies on Nef function, we tested different approaches to restore Nef production in HIV-rtTA. Strategies that involved translation via an EMCV or synthetic internal ribosome entry site (IRES) failed because these elements were incompatible with efficient virus replication. Fusion protein approaches with the FMDV 2A peptide and human ubiquitin were successful and resulted in genetically-stable Nef-expressing HIV-rtTA strains that replicate more efficiently in primary T-cells and human immune system (HIS) mice than Nef-deficient variants, thus confirming the positive effect of Nef on in vivo virus replication. - Highlights: • Different approaches to encode additional proteins in the HIV-1 genome were tested. • IRES translation elements are incompatible with efficient HIV-1 replication. • Ubiquitin and 2A fusion protein approaches allow efficient HIV-1 replication. • Doxycycline-controlled HIV-1 variants that encode all viral proteins were developed. • Nef stimulates HIV-rtTA replication in primary cells and human immune system mice.

  3. Structural basis of evasion of cellular adaptive immunity by HIV-1 Nef

    Energy Technology Data Exchange (ETDEWEB)

    Jia, Xiaofei; Singh, Rajendra; Homann, Stefanie; Yang, Haitao; Guatelli, John; Xiong, Yong (Yale); (VA); (UCSD)

    2012-10-24

    The HIV-1 protein Nef inhibits antigen presentation by class I major histocompatibility complex (MHC-I). We determined the mechanism of this activity by solving the crystal structure of a protein complex comprising Nef, the MHC-I cytoplasmic domain (MHC-I CD) and the {mu}1 subunit of the clathrin adaptor protein complex 1. A ternary, cooperative interaction clamps the MHC-I CD into a narrow binding groove at the Nef-{mu}1 interface, which encompasses the cargo-recognition site of {mu}1 and the proline-rich strand of Nef. The Nef C terminus induces a previously unobserved conformational change in {mu}1, whereas the N terminus binds the Nef core to position it optimally for complex formation. Positively charged patches on {mu}1 recognize acidic clusters in Nef and MHC-I. The structure shows how Nef functions as a clathrin-associated sorting protein to alter the specificity of host membrane trafficking and enable viral evasion of adaptive immunity.

  4. Synthesis and evaluation of orally active small molecule HIV-1 Nef antagonists.

    Science.gov (United States)

    Emert-Sedlak, Lori A; Loughran, H Marie; Shi, Haibin; Kulp, John L; Shu, Sherry T; Zhao, Jielu; Day, Billy W; Wrobel, Jay E; Reitz, Allen B; Smithgall, Thomas E

    2016-03-01

    The HIV-1 Nef accessory factor enhances viral replication and promotes immune system evasion of HIV-infected cells, making it an attractive target for drug discovery. Recently we described a novel class of diphenylpyrazolodiazene compounds that bind directly to Nef in vitro and inhibit Nef-dependent HIV-1 infectivity and replication in cell culture. However, these first-generation Nef antagonists have several structural liabilities, including an azo linkage that led to poor oral bioavailability. The azo group was therefore replaced with either a one- or two-carbon linker. The resulting set of non-azo analogs retained nanomolar binding affinity for Nef by surface plasmon resonance, while inhibiting HIV-1 replication with micromolar potency in cell-based assays without cytotoxicity. Computational docking studies show that these non-azo analogs occupy the same predicted binding site within the HIV-1 Nef dimer interface as the original azo compound. Computational methods also identified a hot spot for inhibitor binding within this site that is defined by conserved HIV-1 Nef residues Asp108, Leu112, and Pro122. Pharmacokinetic evaluation of the non-azo B9 analogs in mice showed that replacement of the azo linkage dramatically enhanced oral bioavailability without substantially affecting plasma half-life or clearance. The improved oral bioavailability of non-azo diphenylpyrazolo Nef antagonists provides a starting point for further drug lead optimization in support of future efficacy testing in animal models of HIV/AIDS.

  5. HIV-1 Nef associates with p22-phox, a component of the NADPH oxidase protein complex.

    Science.gov (United States)

    Salmen, Siham; Colmenares, Melisa; Peterson, Darrel L; Reyes, Elbert; Rosales, Jose D; Berrueta, Lisbeth

    2010-01-01

    Altered neutrophil function may contribute to the development of AIDS during the course of HIV infection. It has been described that Nef, a regulatory protein from HIV, can modulate superoxide production in other cells, therefore altered superoxide production in neutrophils from HIV infected patients, could be secondary to a direct effect of Nef on components of the NADPH oxidase complex. In this work, we describe that Nef, was capable of increasing superoxide production in human neutrophils. Furthermore, a specific association between Nef and p22-phox, a membrane component of the NADPH oxidase complex, was found. We propose that this association may reflect a capability of Nef to modulate by direct association, the enzymatic complex responsible for one of the most efficient innate defense mechanisms in phagocytes, contributing to the pathogenesis of the disease.

  6. Human Immunodeficiency Virus Type 1 Nef Binds to Tumor Suppressor p53 and Protects Cells against p53-Mediated Apoptosis

    OpenAIRE

    2002-01-01

    The nef gene product of human immunodeficiency virus type 1 (HIV-1) is important for the induction of AIDS, and key to its function is its ability to manipulate T-cell function by targeting cellular signal transduction proteins. We reported that Nef coprecipitates a multiprotein complex from cells which contains tumor suppressor protein p53. We now show that Nef interacts directly with p53. Binding assays showed that an N-terminal, 57-residue fragment of Nef (Nef 1-57) contains the p53-bindin...

  7. HIV-1 Nef interaction influences the ATP-binding site of the Src-family kinase, Hck

    Directory of Open Access Journals (Sweden)

    Pene-Dumitrescu Teodora

    2012-03-01

    Full Text Available Abstract Background Nef is an HIV-1 accessory protein essential for viral replication and AIDS progression. Nef interacts with a multitude of host cell signaling partners, including members of the Src kinase family. Nef preferentially activates Hck, a Src-family kinase (SFK strongly expressed in macrophages and other HIV target cells, by binding to its regulatory SH3 domain. Recently, we identified a series of kinase inhibitors that preferentially inhibit Hck in the presence of Nef. These compounds also block Nef-dependent HIV replication, validating the Nef-SFK signaling pathway as an antiretroviral drug target. Our findings also suggested that by binding to the Hck SH3 domain, Nef indirectly affects the conformation of the kinase active site to favor inhibitor association. Results To test this hypothesis, we engineered a "gatekeeper" mutant of Hck with enhanced sensitivity to the pyrazolopyrimidine tyrosine kinase inhibitor, NaPP1. We also modified the RT loop of the Hck SH3 domain to enhance interaction of the kinase with Nef. This modification stabilized Nef:Hck interaction in solution-based kinase assays, as a way to mimic the more stable association that likely occurs at cellular membranes. Introduction of the modified RT loop rendered Hck remarkably more sensitive to activation by Nef, and led to a significant decrease in the Km for ATP as well as enhanced inhibitor potency. Conclusions These observations suggest that stable interaction with Nef may induce Src-family kinase active site conformations amenable to selective inhibitor targeting.

  8. Expression, intracellular targeting and purification of HIV Nef variants in tobacco cells

    Directory of Open Access Journals (Sweden)

    Baschieri Selene

    2007-02-01

    Full Text Available Abstract Background Plants may represent excellent alternatives to classical heterologous protein expression systems, especially for the production of biopharmaceuticals and vaccine components. Modern vaccines are becoming increasingly complex, with the incorporation of multiple antigens. Approaches towards developing an HIV vaccine appear to confirm this, with a combination of candidate antigens. Among these, HIV-Nef is considered a promising target for vaccine development because immune responses directed against this viral protein could help to control the initial steps of viral infection and to reduce viral loads and spreading. Two isoforms of Nef protein can be found in cells: a full-length N-terminal myristoylated form (p27, 27 kDa and a truncated form (p25, 25 kDa. Here we report the expression and purification of HIV Nef from transgenic tobacco. Results We designed constructs to direct the expression of p25 and p27 Nef to either the cytosol or the secretory pathway. We tested these constructs by transient expression in tobacco protoplasts. Cytosolic Nef polypeptides are correctly synthesised and are stable. The same is not true for Nef polypeptides targeted to the secretory pathway by virtue of a signal peptide. We therefore generated transgenic plants expressing cytosolic, full length or truncated Nef. Expression levels were variable, but in some lines they averaged 0.7% of total soluble proteins. Hexahistidine-tagged Nef was easily purified from transgenic tissue in a one-step procedure. Conclusion We have shown that transient expression can help to rapidly determine the best cellular compartment for accumulation of a recombinant protein. We have successfully expressed HIV Nef polypeptides in the cytosol of transgenic tobacco plants. The proteins can easily be purified from transgenic tissue.

  9. HIV Type 1 Nef Is Released from Infected Cells in CD45+ Microvesicles and Is Present in the Plasma of HIV-Infected Individuals

    Science.gov (United States)

    Raymond, A.D.; Campbell-Sims, T.C.; Khan, M.; Lang, M.; Huang, M.B.; Bond, V.C.

    2011-01-01

    Abstract HIV-1 Nef has been demonstrated to be integral for viral persistence, infectivity, and the acceleration of disease pathogenesis (AIDS) in humans. Nef has also been detected in the plasma of HIV-infected individuals and is released from infected cells. The form in which Nef is released from infected cells is unknown. However, Nef is a myristoylated protein and has been shown to interact with the intracellular vesicular trafficking network. Here we show that Nef is released in CD45-containing microvesicles. This microvesicular Nef (mvNef) is detected in the plasma of HIV-infected individuals at relatively high concentrations (10 ng/ml). It is also present in tissue culture supernatants of Jurkat cells infected with HIVMN. Interestingly, plasma mvNef levels in HIV+ patients did not significantly correlate with viral load or CD4 count. Microvesicular Nef levels persisted in the plasma of HIV-infected individuals despite the use of antiretroviral therapy, even in individuals with undetectable viral loads. Using cell lines, we found Nef microvesicles induce apoptosis in Jurkat T-lymphocytes but had no observed effect on the U937 monocytic cell line. Given the large amount of mvNef present in the plasma of HIV-infected individuals, the apoptotic effect of mvNef on T cells, and the observed functions of extracellular soluble Nef in vitro, it seems likely that in vivo mvNef may play a significant role in the pathogenesis of AIDS. PMID:20964480

  10. Lipidomic dataset of plasma from patients infected with wild type and nef-deficient HIV-1 strain

    Directory of Open Access Journals (Sweden)

    Peter Meikle

    2016-03-01

    Full Text Available Previous in vitro and in vivo studies demonstrated that HIV protein nef plays a key role in impairing cellular and systemic cholesterol metabolism in HIV disease, but clinical support for these findings is lacking. Here we present the data of comparative lipidomic analysis (330 lipid species of plasma samples from HIV-negative subjects, patients infected with WT HIV-1 strain and patients infected with nef-deficient strain of HIV-1. We determine which effects of HIV on plasma lipidome are explained by the presence of nef. The data can be used to evaluate cardiovascular risk in HIV disease and to assess the role of nef in HIV-induced disturbances in systemic lipid metabolism. The full impact of nef deficiency on lipid and lipoprotein metabolism in HIV-infected patients is presented in the accompanying study “Lipid Metabolism in Patients Infected with Nef-deficient HIV-1 Strain” [1].

  11. High levels of anti-Nef antibodies may prevent AIDS disease progression in vertically HIV-1-infected infants

    Directory of Open Access Journals (Sweden)

    Guillermo Corró

    2014-02-01

    Full Text Available Introduction: HIV-1-associated CD4+ T-cell depletion is a consequence of uninfected cell death. Nef is one of the viral factors that trigger apoptosis on bystander cells, though the plasma Nef levels do not correlate with Th lymphocytes counts. The aim of our study was to evaluate whether anti-Nef antibodies were involved in paediatric AIDS development and whether they can prevent the CD4+ T-cell depletion in vertically infected children. Methods: Two hundred and seventy three HIV-1 vertically infected children seen at Garrahan Paediatric Hospital were randomly included in the study, adding 13 selected cases: seven LTNP (long-term non-progressors and six RP (rapid progressors children (ntotal=286. Specific anti-HIV-1-Nef antibodies were titrated by indirect ELISA and compared between groups. The plasma blocking effect on Nef-dependent cytotoxicity was evaluated in Jurkat cells using recombinant Nef as apoptotic stimulus and patient plasmas as blockers, measuring the apoptotic levels using Annexin-V stain and flow cytometry. Results: Only 63.4% of the patients had specific anti-Nef antibodies, and the levels of anti-Nef antibodies found in the selected LTNPs plasmas were always significantly higher (p=1.55×10−4 than those in RPs or general HIV-1+ paediatric populations. The LTNPs’ plasma had a strong inhibitory effect on Nef-dependent cytotoxicity even at high dilutions, while RP plasmas had little or no effect on Nef-induced apoptosis. Discussion and conclusions: High anti-Nef antibody levels are associated and predict slow or non-progression to AIDS in vertically HIV-1-infected children. They could be an efficient tool in preventing Nef-associated bystander effect, preserving CD4+ T-cells and the immune function in the context of paediatric HIV-1 infection.

  12. HIV-1 Nef impairs key functional activities in human macrophages through CD36 downregulation.

    Directory of Open Access Journals (Sweden)

    Eleonora Olivetta

    Full Text Available Monocytes and macrophages utilize the class A and B scavenger receptors to recognize and perform phagocytosis of invading microbes before a pathogen-specific immune response is generated. HIV-1 Nef protein affects the innate immune system impairing oxidative burst response and phagocytic capacity of macrophages. Our data show that exogenous recombinant myristoylated Nef protein induces a marked CD36 downregulation in monocytes from Peripheral Blood Mononuclear Cells, in Monocyte-Derived Macrophages (MDMs differentiated by cytokines and in MDMs contained in a mixed culture obtained expanding PBMCs under Human Erythroid Massive Amplification condition. Under the latter culture condition we identify three main populations after 6 days of expansion: lymphocytes (37.8 ± 14.7%, erythroblasts (46.7±6.1% and MDMs (15.7 ± 7.5%. The Nef addition to the cell culture significantly downregulates CD36 expression in MDMs, but not in erythroid cells. Furthermore, CD36 inhibition is highly specific since it does not modify the expression levels of other MDM markers such as CD14, CD11c, CD86, CD68, CD206, Toll-like Receptor 2 and Toll-like Receptor 4. Similar results were obtained in MDMs infected with VSV-G pseudotyped HIV-1-expressing Nef. The reduced CD36 membrane expression is associated with decrease of correspondent CD36 mRNA transcript. Furthermore, Nef-induced CD36 downregulation is linked to both impaired scavenger activity with reduced capability to take up oxidized lipoproteins and to significant decreased phagocytosis of fluorescent beads and GFP-expressing Salmonella tiphymurium. In addition we observed that Nef induces TNF-α release in MDMs. Although these data suggest a possible involvement of TNF-α in mediating Nef activity, our results exclude a possible relationship between Nef-induced TNF-α release and Nef-mediated CD36 downregulation. The present work shows that HIV-1 Nef protein may have a role in the strategies elaborated by HIV-1 to

  13. Exogenous HIV-1 Nef upsets the IFN-γ-induced impairment of human intestinal epithelial integrity.

    Directory of Open Access Journals (Sweden)

    Maria Giovanna Quaranta

    Full Text Available BACKGROUND: The mucosal tissues play a central role in the transmission of HIV-1 infection as well as in the pathogenesis of AIDS. Despite several clinical studies reported intestinal dysfunction during HIV infection, the mechanisms underlying HIV-induced impairments of mucosal epithelial barrier are still unclear. It has been postulated that HIV-1 alters enterocytic function and HIV-1 proteins have been detected in several cell types of the intestinal mucosa. In the present study, we analyzed the effect of the accessory HIV-1 Nef protein on human epithelial cell line. METHODOLOGY/PRINCIPAL FINDINGS: We used unstimulated or IFN-γ-stimulated Caco-2 cells, as a model for homeostatic and inflamed gastrointestinal tracts, respectively. We investigated the effect of exogenous recombinant Nef on monolayer integrity analyzing its uptake, transepithelial electrical resistance, permeability to FITC-dextran and the expression of tight junction proteins. Moreover, we measured the induction of proinflammatory mediators. Exogenous Nef was taken up by Caco-2 cells, increased intestinal epithelial permeability and upset the IFN-γ-induced reduction of transepithelial resistance, interfering with tight junction protein expression. Moreover, Nef inhibited IFN-γ-induced apoptosis and up-regulated TNF-α, IL-6 and MIP-3α production by Caco-2 cells while down-regulated IL-10 production. The simultaneous exposure of Caco-2 cells to Nef and IFN-γ did not affect cytokine secretion respect to untreated cells. Finally, we found that Nef counteracted the IFN-γ induced arachidonic acid cascade. CONCLUSION/SIGNIFICANCE: Our findings suggest that exogenous Nef, perturbing the IFN-γ-induced impairment of intestinal epithelial cells, could prolong cell survival, thus allowing for accumulation of viral particles. Our results may improve the understanding of AIDS pathogenesis, supporting the discovery of new therapeutic interventions.

  14. Trans-cellular introduction of HIV-1 protein Nef induces pathogenic response in Caenorhabditis elegans.

    Directory of Open Access Journals (Sweden)

    Aamir Nazir

    Full Text Available BACKGROUND: Caenorhabditis elegans has emerged as a very powerful model for studying the host pathogen interactions. Despite the absence of a naturally occurring viral infection for C. elegans, the model is now being exploited experimentally to study the basic aspects of virus-host interplay. The data generated from recent studies suggests that the virus that infects mammalian cells does infect, replicate and accumulate in C. elegans. METHODOLOGY/PRINCIPAL FINDINGS: We took advantage of the easy-to-achieve protein introduction in C. elegans and employing the methodology, we administered HIV-1 protein Nef into live worms. Nef is known to be an important protein for exacerbating HIV-1 pathogenesis in host by enhancing viral replication. The deletion of nef from the viral genome has been reported to inhibit its replication in the host, thereby leading to delayed pathogenesis. Our studies, employing Nef introduction into C. elegans, led to creation of an in-vivo model that allowed us to study, whether or not, the protein induces effect in the whole organism. We observed a marked lipodystrophy, effect on neuromuscular function, impaired fertility and reduced longevity in the worms exposed to Nef. The observed effects resemble to those observed in Nef transgenic mice and most interestingly the effects also relate to some of the pathogenic aspects exhibited by human AIDS patients. CONCLUSIONS/SIGNIFICANCE: Our studies underline the importance of this in vivo model for studying the interactions of Nef with host proteins, which could further be used for identifying possible inhibitors of such interactions.

  15. Vegetation recovery in tidal marshes reveals critical slowing down under increased inundation

    Science.gov (United States)

    van Belzen, Jim; van de Koppel, Johan; Kirwan, Matthew L.; van der Wal, Daphne; Herman, Peter M. J.; Dakos, Vasilis; Kéfi, Sonia; Scheffer, Marten; Guntenspergen, Glenn R.; Bouma, Tjeerd J.

    2017-06-01

    A declining rate of recovery following disturbance has been proposed as an important early warning for impending tipping points in complex systems. Despite extensive theoretical and laboratory studies, this `critical slowing down' remains largely untested in the complex settings of real-world ecosystems. Here, we provide both observational and experimental support of critical slowing down along natural stress gradients in tidal marsh ecosystems. Time series of aerial images of European marsh development reveal a consistent lengthening of recovery time as inundation stress increases. We corroborate this finding with transplantation experiments in European and North American tidal marshes. In particular, our results emphasize the power of direct observational or experimental measures of recovery over indirect statistical signatures, such as spatial variance or autocorrelation. Our results indicate that the phenomenon of critical slowing down can provide a powerful tool to probe the resilience of natural ecosystems.

  16. Network modularity reveals critical scales for connectivity in ecology and evolution.

    Science.gov (United States)

    Fletcher, Robert J; Revell, Andre; Reichert, Brian E; Kitchens, Wiley M; Dixon, Jeremy D; Austin, James D

    2013-01-01

    For nearly a century, biologists have emphasized the profound importance of spatial scale for ecology, evolution and conservation. Nonetheless, objectively identifying critical scales has proven incredibly challenging. Here we extend new techniques from physics and social sciences that estimate modularity on networks to identify critical scales for movement and gene flow in animals. Using four species that vary widely in dispersal ability and include both mark-recapture and population genetic data, we identify significant modularity in three species, two of which cannot be explained by geographic distance alone. Importantly, the inclusion of modularity in connectivity and population viability assessments alters conclusions regarding patch importance to connectivity and suggests higher metapopulation viability than when ignoring this hidden spatial scale. We argue that network modularity reveals critical meso-scales that are probably common in populations, providing a powerful means of identifying fundamental scales for biology and for conservation strategies aimed at recovering imperilled species.

  17. Network modularity reveals critical scales for connectivity in ecology and evolution

    Science.gov (United States)

    Fletcher, Robert J.; Revell, Andre; Reichert, Brian E.; Kitchens, Wiley M.; Dixon, J.; Austin, James D.

    2013-01-01

    For nearly a century, biologists have emphasized the profound importance of spatial scale for ecology, evolution and conservation. Nonetheless, objectively identifying critical scales has proven incredibly challenging. Here we extend new techniques from physics and social sciences that estimate modularity on networks to identify critical scales for movement and gene flow in animals. Using four species that vary widely in dispersal ability and include both mark-recapture and population genetic data, we identify significant modularity in three species, two of which cannot be explained by geographic distance alone. Importantly, the inclusion of modularity in connectivity and population viability assessments alters conclusions regarding patch importance to connectivity and suggests higher metapopulation viability than when ignoring this hidden spatial scale. We argue that network modularity reveals critical meso-scales that are probably common in populations, providing a powerful means of identifying fundamental scales for biology and for conservation strategies aimed at recovering imperilled species.

  18. SIVsm Tat, Rev, and Nef1: functional characteristics of r-GV internalization on isotypes, cytokines, and intracellular degradation

    Directory of Open Access Journals (Sweden)

    Stuart Elizabeth S

    2010-07-01

    Full Text Available Abstract Background Recombinant gas vesicles (r-GV from Halobacterium sp. strain SD109 expressing cassettes with different SIVsm inserts, have potential utility as an effective antigen display system for immunogen testing in vivo and for initial epitope assessments in vitro. Previous mouse model studies demonstrated immunization with r-GV expressing selected exogenous sequences elicited a prolonged immune response. Here we tested segments from three SIVsm genes (tat, rev, and nef each surface displayed by r-GV. As with HIV, for SIVsm the proteins encoded by tat, rev and nef respectively serve critical and diverse functions: effects on efficient viral RNA polymerase II transcription, regulation of viral gene expression and effects on specific signaling functions through the assembly of multiprotein complexes. Humoral responses to r-GVTat, Rev or Nef1 elicited in vivo, associated changes in selected cell cytokine production following r-GV internalization, and the capacity of J774A.1 macrophage cells to degrade these internalized display/delivery particles in vitro were examined. Results The in vivo studies involving r-GV immunizations and in vitro studies of r-GV uptake by J774A.1 macrophages demonstrated: (i tests for antibody isotypes in immunized mice sera showed activation and re-stimulation of memory B cells, (ii during long term immune response to the epitopes, primarily the IgG1 isotype was produced, (iii in vitro, macrophage degradation of r-GV containing different SIVsm inserts occurred over a period of days resulting in an inherent slow breakdown and degradation of the SIVsm peptide inserts, (iv vesicle specific GvpC, a larger protein, degraded more slowly than the recombinant peptide inserts and (v in vitro uptake and degradation of the r-GV populations tested was associated with SIVsm insert specific patterns for cytokines IL-10, IL-12 and IL-18. Conclusions Together these findings provide new information underscoring r-GV potential

  19. [Variability of the HIV-1 nef regulatory gene and its association with different HIV stages].

    Science.gov (United States)

    Ryzhov, K A; Matsevich, G R; Gol'tsov, V A; Lariukova, T A; Zverev, V V

    2004-01-01

    Biological properties of HIV-1 laboratory strains and isolates were studies and compared with research results of the nef gene fragment obtained from their proviral DNA and RNA as well as from RNA and HIV-1 DNA isolated immediately from the blood of patients at early HIV stages. The electrophoresis pattern as well as the results of determination of nucleotide sequences showed that the HVI-1 RNA nef gene of rapid/high laboratory strains of HIV isolates and RNA obtained from patients at late infection stages had a 135-nucleotide inner deletion. The phenomenon can be regarded is proof of that nef gene structure, if violated, causes an enhanced virulence of and an intensified multiplication of the virus (according to laboratory markers) observed at late HIV stages, which triggers, at least in a number of cases, the infection aggravation.

  20. Revealing the transformatory moment of learning technology: the place of critical social theory

    Directory of Open Access Journals (Sweden)

    Richard Hall

    2011-12-01

    Full Text Available In revealing the transformatory moment of learning technology, a holistic andongoing critique that is related to a range of socio-cultural factors is required.This critique further uncovers the social relations that frame the place of learningtechnology in bourgeois capitalist work and the development of the knowledgeeconomy. This article argues that critical social theory, and the social and powerrelationships that it highlights, is central to the uncovering of this revelation. Asa result of a holistic, or totalising, analysis based upon the political economy oftechnology in education, a framework that enables educators to examine its educationalvalue is uncovered. Iterating and re-producing this framework works asa constant critique of educational practice within capitalism, and critiques deterministicor hegemonic views of technology-as-progress. As a result, this articleseeks to demonstrate how critical social theory might enable educators to analyselearning technology as historically situated, and thereby to use critique to revealand produce practical alternatives to sociocultural problems.

  1. HIV-Associated Pathogenesis: The Role of HIV-1 Nef Gene

    Institute of Scientific and Technical Information of China (English)

    Xiaoning Xu; Hao Wu

    2006-01-01

    Since the discovery of HIV more than two decades ago, scientific progress has been impressive and has dramatically advanced our understanding of HIV infection, from molecular through cellular and then on to systemic pathogenesis,such as the functions of HIV-1 nef gene. However, there is still much to be learned before we fully understand how the host interacts with the virus at molecular level and how immune responses correlate with protection from disease. This review describes our current knowledge of HIV/AIDS with the reference of HIV nef functions and its role in HIV-1 mediated pathogenesis.

  2. HIV-1 Nef protein in the nucleus influences adipogenesis as well as viral transcription through the peroxisome proliferator-activated receptors.

    Science.gov (United States)

    Otake, Kaori; Omoto, Shinya; Yamamoto, Takuya; Okuyama, Harumi; Okada, Hidechika; Okada, Noriko; Kawai, Masahiro; Saksena, Nitin K; Fujii, Yoichi R

    2004-01-23

    Although the HIV-1 Nef protein (27 kDa) localizes primarily in cytoplasm, there is considerable evidence suggesting its occasional localization in the nucleus. Nef is known to play an important role in transcriptional events and viral replication, but the actual target of Nef in the nucleus remains to be identified. To examine the functional roles of Nef in the nucleus and its possible interactions with other unknown factors in the nucleus. High-density microarray analysis was used to screen directly the unique functions of Nef on host gene transcription. The nuclear localization of Nef and its effects on the expression of peroxisome proliferator-activated receptors (PPAR) was examined using PPAR promoter/reporter assay and immunoblotting. A long terminal repeat/reporter assay was used to investigated the effects of Nef and PPAR on viral transcription. Nef in the nucleus suppressed PPAR gamma expression and reduced fatty acid levels in human T and macrophage cell lines. Expression of Nef or PPAR suppressed viral replication; the effect of PPAR gamma or retinoid X receptor-alpha on viral replication were reduced by coexpression of Nef in MT(-)4 T cells. Nef may be involved in both viral replication and the wasting syndrome associated with AIDS.

  3. Monoclonal antibodies against a synthetic peptide from human immunodeficiency virus type 1 Nef protein

    DEFF Research Database (Denmark)

    Steinaa, L; Wulff, A M; Saermark, T

    1994-01-01

    Monoclonal antibodies against a synthetic peptide (aa 138-152) from HIV-1 Nef protein were produced and characterized. Three hybridoma lines producing monoclonal antibodies (MAbs) against the synthetic peptide were generated by fusion between P3-X63 Ag8.653 myeloma cells and BALB/c splenocytes fr...

  4. HIV-1 Protein Nef Inhibits Activity of ATP-binding Cassette Transporter A1 by Targeting Endoplasmic Reticulum Chaperone Calnexin*

    Science.gov (United States)

    Jennelle, Lucas; Hunegnaw, Ruth; Dubrovsky, Larisa; Pushkarsky, Tatiana; Fitzgerald, Michael L.; Sviridov, Dmitri; Popratiloff, Anastas; Brichacek, Beda; Bukrinsky, Michael

    2014-01-01

    HIV-infected patients are at increased risk of developing atherosclerosis, in part due to an altered high density lipoprotein profile exacerbated by down-modulation and impairment of ATP-binding cassette transporter A1 (ABCA1) activity by the HIV-1 protein Nef. However, the mechanisms of this Nef effect remain unknown. Here, we show that Nef interacts with an endoplasmic reticulum chaperone calnexin, which regulates folding and maturation of glycosylated proteins. Nef disrupted interaction between calnexin and ABCA1 but increased affinity and enhanced interaction of calnexin with HIV-1 gp160. The Nef mutant that did not bind to calnexin did not affect the calnexin-ABCA1 interaction. Interaction with calnexin was essential for functionality of ABCA1, as knockdown of calnexin blocked the ABCA1 exit from the endoplasmic reticulum, reduced ABCA1 abundance, and inhibited cholesterol efflux; the same effects were observed after Nef overexpression. However, the effects of calnexin knockdown and Nef on cholesterol efflux were not additive; in fact, the combined effect of these two factors together did not differ significantly from the effect of calnexin knockdown alone. Interestingly, gp160 and ABCA1 interacted with calnexin differently; although gp160 binding to calnexin was dependent on glycosylation, glycosylation was of little importance for the interaction between ABCA1 and calnexin. Thus, Nef regulates the activity of calnexin to stimulate its interaction with gp160 at the expense of ABCA1. This study identifies a mechanism for Nef-dependent inactivation of ABCA1 and dysregulation of cholesterol metabolism. PMID:25170080

  5. Association between Nef-specific CD8 T-cell responses and disease progression in HIV-1 subtype B infection

    Institute of Scientific and Technical Information of China (English)

    JIAO Yang; LI Tai-sheng; XIE Jing; HAN Yang; QIU Zhi-feng; ZUO Ling-yan; Thomas Mourez; WANG Ai-xia

    2006-01-01

    Background The correlation between HIV-1 Nef-specific CD8 T-cell responses and markers of HIV-1 disease progression still remains unclear. This study analysed and compared the role of HIV-1 Nef-specific CD8 T-cell responses in patients with different disease status.Methods Two groups of patients with HIV-1 subtype B infection were selected according to CD4 count and clinical manifestations: long-term nonprogresssors (LTNPs, n = 20) and advanced progressors (Aps, CD4 count <500 cells/μ1, n = 34). Nef-specific CD8 T-cell responses were studied by interferon- γ ELISpot assay against 3 pools of HIV-Nef peptides.Results Nef-specific CD8 T-cell responses did not correlate with viral load or CD4 count in all patients and no significant differences were found in the magnitude of Nef-specific CD8 T-cell responses between groups LTNPs and Aps (670 SFC/106 peripheral blood mononuclear cells vs 1107 SFC/106 peripheral blood mononuclear cells,P = 0.255). Further comparisons showed that there were also no significant correlations observed in group LTNPs,but Nef-specific CD8 T cells correlated negatively with viral load (r = -0.397, P = 0.020) and positively with CD4 count (r = 0.364, P = 0.034) in group Aps.Conclusion These data suggest that different correlation patterns between Nef-specific CD8 T-cell responses and disease progression exist in LTNPs and Aps. Although a negative association was observed with concurrent plasma HIV RNA in Aps, Nef-specific CD8 T-cell responses might fail to play a protective role in different stages of HIV- 1 infection.

  6. A homology model of Xyloglucan Xylosyltransferase 2 reveals critical amino acids involved in substrate binding.

    Science.gov (United States)

    Culbertson, Alan T; Tietze, Alesia A; Tietze, Daniel; Chou, Yi-Hsiang; Smith, Adrienne L; Young, Zachary T; Zabotina, Olga A

    2016-09-01

    In dicotyledonous plants, xyloglucan (XyG) is the most abundant hemicellulose of the primary cell wall. The enzymes involved in XyG biosynthesis have been identified through reverse-genetics and activity was characterized by heterologous expression. Currently, there is no information on the atomic structures or amino acids involved in activity or substrate binding of any of the Golgi-localized XyG biosynthetic enzymes. A homology model of the xyloglucan xylosyltransferase 2 (XXT2) catalytic domain was built on the basis of the crystal structure of A64Rp. Molecular dynamics simulations revealed that the homology model retains the glycosyltransferase (GT)-A fold of the template structure used to build the homology model indicating that XXT2 likely has a GT-A fold. According to the XXT2 homology model, six amino acids (Phe204, Lys207, Asp228, Ser229, Asp230, His378) were selected and their contribution in catalytic activity was investigated. Site-directed mutagenesis studies show that Asp228, Asp230 and His378 are critical for XXT2 activity and are predicted to be involved in coordination of manganese ion. Lys207 was also found to be critical for protein activity and the homology model indicates a critical role in substrate binding. Additionally, Phe204 mutants have less of an impact on XXT2 activity with the largest effect when replaced with a polar residue. This is the first study that investigates the amino acids involved in substrate binding of the XyG-synthesizing xylosyltransferases and contributes to the understanding of the mechanisms of polysaccharide-synthesizing GTs and XyG biosynthesis. © The Author 2016. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  7. Microsatellite variability reveals high genetic diversity and low genetic differentiation in a critical giant panda population

    Institute of Scientific and Technical Information of China (English)

    Jiandong YANG; Zhihe ZHANG; Fujun SHEN; Xuyu YANG; Liang ZHANG; Limin CHEN; Wenping ZHANG; Qing ZHU; Rong HOU

    2011-01-01

    Understanding present patterns of genetic diversity is critical in order to design effective conservation and management strategies for endangered species.Tangjiahe Nature Reserve (NR) is one of the most important national reserves for giant pandas Ailuropoda melanoleuca in China.Previous studies have shown that giant pandas in Tangjiahe NR may be threatened by population decline and fragmentation.Here we used 10 microsatellite DNA markers to assess the genetic variability in the Tangjiahe population.The results indicate a low level of genetic differentiation between the Hongshihe and Motianling subpopulations in the reserve.Assignment tests using the Bayesian clustering method in STRUCTURE identified one genetic cluster from 42 individuals of the two subpopulations.All individuals from the same subpopulation were assigned to one cluster.This indicates high gene flow between subpopulations.F statistic analyses revealed a low Fls-value of 0.024 in the total population and implies a randomly mating population in Tangjiahe NR.Additionally,our data show a high level of genetic diversity for the Tangjiahe population.Mean allele number (A),Allelic richness (AR) and mean expected heterozygosity (HE) for the Tangiiahe population was 5.9,5.173 and 0.703,respectively.This wild giant panda population can be restored through concerted effort [Current Zoology 57 (6):717-724,2011].

  8. Microsatellite variability reveals high genetic diversity and low genetic differentiation in a critical giant panda population

    Directory of Open Access Journals (Sweden)

    Jiandong YANG, Zhihe ZHANG, Fujun SHEN, Xuyu YANG, Liang ZHANG, Limin CHEN, Wenping ZHANG, Qing ZHU, Rong HOU

    2011-12-01

    Full Text Available Understanding present patterns of genetic diversity is critical in order to design effective conservation and management strategies for endangered species. Tangjiahe Nature Reserve (NR is one of the most important national reserves for giant pandas Ailuropoda melanoleuca in China. Previous studies have shown that giant pandas in Tangjiahe NR may be threatened by population decline and fragmentation. Here we used 10 microsatellite DNA markers to assess the genetic variability in the Tangjiahe population. The results indicate a low level of genetic differentiation between the Hongshihe and Motianling subpopulations in the reserve. Assignment tests using the Bayesian clustering method in STRUCTURE identified one genetic cluster from 42 individuals of the two subpopulations. All individuals from the same subpopulation were assigned to one cluster. This indicates high gene flow between subpopulations. F statistic analyses revealed a low FIS-value of 0.024 in the total population and implies a randomly mating population in Tangjiahe NR. Additionally, our data show a high level of genetic diversity for the Tangjiahe population. Mean allele number (A, Allelic richness (AR and mean expected heterozygosity (HE for the Tangjiahe population was 5.9, 5.173 and 0.703, respectively. This wild giant panda population can be restored through concerted effort [Current Zoology 57 (6: 717–724, 2011].

  9. Conditional Knockout in Mice Reveals the Critical Roles of Ppp2ca in Epidermis Development

    Directory of Open Access Journals (Sweden)

    Chao Fang

    2016-05-01

    Full Text Available The epidermis is an important tissue in Homo sapines and other animals, and an abnormal epidermis will cause many diseases. Phosphatase 2A (PP2A is an important serine and threonine phosphatase. The α isoform of the PP2A catalytic subunit (Ppp2ca gene encoding PP2Acα is critical for cell proliferation, growth, metabolism and tumorigenesis. However, to date, no study has revealed its roles in epidermis development. To specifically investigate the roles of PP2Acα in epidermis development, we first generated Ppp2caflox/flox transgenic mice, and conditionally knocked out Ppp2ca in the epidermis driven by Krt14-Cre. Our study showed that Ppp2caflox/flox; Krt14-Cre mice had significant hair loss. In addition, histological analyses showed that the morphogenesis and hair regeneration cycle of hair follicles were disrupted in these mice. Moreover, Ppp2caflox/flox; Krt14-Cre mice had smaller size, melanin deposition and hyperproliferation at the base of the claws. Accordingly, our study demonstrates that PP2Acα plays important roles in both hair follicle and epidermis development. Additionally, the Ppp2caflox/flox mice generated in this study can serve as a useful transgene model to study the roles of PP2Acα in other developmental processes and diseases.

  10. Cross-clade conservation of HIV type 1 Nef immunodominant regions recognized by CD8+ T cells of HIV type 1 CRF02_AG-infected Ivorian (West Africa).

    Science.gov (United States)

    Inwoley, André; Recordon-Pinson, Patricia; Dupuis, Marion; Gaston, Jessintha; Genête, Mathieu; Minga, Albert; Letourneur, Franck; Rouet, François; Choppin, Jeannine; Fleury, Hervé; Guillet, Jean-Gérard; Andrieu, Muriel

    2005-07-01

    Most HIV vaccine trials in the world are conducted with clade B while most circulating viral strains in Africa are non-B subtypes. We determined whether CD8+ T cells from HIV-1 intersubtype CRF02_AG-infected Ivorian individuals were able to recognize clade B epitopes. CD8+ T cell responses of nine HIV-1 intersubtype CRF02_AG-infected Ivorian patients and nine HIV-1 subtype B-infected French patients were studied using pools of HIV-1 clade B peptides (110 well-defined HIV CD8+ T cell epitopes) in an ELISPOT IFN-gamma assay. There was no difference in the number of recognized peptide pools between Ivorian and French cohorts (mean of four pools in both cases). Ivorian individuals had generated CD8+ T cell responses cross-reactive against HIV-1 subtype B and some individual peptides had been identified. Furthermore, sequence analysis of nef HIV genes of the Ivorian patients and nef cloning in two patients revealed very few variations between HIV- 1 intersubtype CRF02_AG and subtype B in nef immunodominant regions included in HIV clade B lipopeptide vaccines, currently tested in France.

  11. Nef alleles from all major HIV-1 clades activate Src-family kinases and enhance HIV-1 replication in an inhibitor-sensitive manner.

    Directory of Open Access Journals (Sweden)

    Purushottam S Narute

    Full Text Available The HIV-1 accessory factor Nef is essential for high-titer viral replication and AIDS progression. Nef function requires interaction with many host cell proteins, including specific members of the Src kinase family. Here we explored whether Src-family kinase activation is a conserved property of Nef alleles from a wide range of primary HIV-1 isolates and their sensitivity to selective pharmacological inhibitors. Representative Nef proteins from the major HIV-1 subtypes A1, A2, B, C, F1, F2, G, H, J and K strongly activated Hck and Lyn as well as c-Src to a lesser extent, demonstrating for the first time that Src-family kinase activation is a highly conserved property of primary M-group HIV-1 Nef isolates. Recently, we identified 4-amino substituted diphenylfuropyrimidines (DFPs that selectively inhibit Nef-dependent activation of Src-family kinases as well as HIV replication. To determine whether DFP compounds exhibit broad-spectrum Nef-dependent antiretroviral activity against HIV-1, we first constructed chimeric forms of the HIV-1 strain NL4-3 expressing each of the primary Nef alleles. The infectivity and replication of these Nef chimeras was indistinguishable from that of wild-type virus in two distinct cell lines (U87MG astroglial cells and CEM-T4 lymphoblasts. Importantly, the 4-aminopropanol and 4-aminobutanol derivatives of DFP potently inhibited the replication of all chimeric forms of HIV-1 in both U87MG and CEM-T4 cells in a Nef-dependent manner. The antiretroviral effects of these compounds correlated with inhibition of Nef-dependent activation of endogenous Src-family kinases in the HIV-infected cells. Our results demonstrate that the activation of Hck, Lyn and c-Src by Nef is highly conserved among all major clades of HIV-1 and that selective targeting of this pathway uniformly inhibits HIV-1 replication.

  12. Human Immunodeficiency Virus Type 1 Nef Inhibits Autophagy through Transcription Factor EB Sequestration.

    Directory of Open Access Journals (Sweden)

    Grant R Campbell

    2015-06-01

    Full Text Available HIV Nef acts as an anti-autophagic maturation factor through interaction with beclin-1 (BECN1. We report that exposure of macrophages to infectious or non-infectious purified HIV induces toll-like receptor 8 (TLR8 and BECN1 dependent dephosphorylation and nuclear translocation of TFEB and that this correlates with an increase in autophagy markers. RNA interference for ATG13, TFEB, TLR8, or BECN1 inhibits this HIV-induced autophagy. However, once HIV establishes a productive infection, TFEB phosphorylation and cytoplasmic sequestration are increased resulting in decreased autophagy markers. Moreover, by 7 d post-infection, autophagy levels are similar to mock infected controls. Conversely, although Nef deleted HIV similarly induces TFEB dephosphorylation and nuclear localization, and increases autophagy, these levels remain elevated during continued productive infection. Thus, the interaction between HIV and TLR8 serves as a signal for autophagy induction that is dependent upon the dephosphorylation and nuclear translocation of TFEB. During permissive infection, Nef binds BECN1 resulting in mammalian target of rapamycin (MTOR activation, TFEB phosphorylation and cytosolic sequestration, and the inhibition of autophagy. To our knowledge, this is the first report of a virus modulating TFEB localization and helps to explain how HIV modulates autophagy to promote its own replication and cell survival.

  13. HIV-1-infection of T lymphocytes and macrophages affects their migration via Nef

    Directory of Open Access Journals (Sweden)

    Christel eVérollet

    2015-10-01

    Full Text Available The human immunodeficiency virus (HIV-1 disseminates in the body and is found in several organs and tissues. While HIV-1 mainly targets both CD4+ T lymphocytes and macrophages, it has contrasting effects between these cell populations. HIV-1 infection namely reduces the viability of CD4+ T cells, whereas infected macrophages are long-lived. In addition, the migration of T cells is reduced by the infection, while HIV-1 differentially modulates the migration modes of macrophages. In 2-dimensions (2D assays, infected macrophages are less motile compared to the control counterparts. In 3D environments, macrophages use two migration modes that are dependent on the matrix architecture: amoeboid and mesenchymal migration. HIV-1 infected macrophages exhibit a reduced amoeboid migration but an enhanced mesenchymal migration, via the viral protein Nef. Indeed, the mesenchymal migration involves podosomes, and Nef stabilizes these cell structures through the activation of the tyrosine kinase Hck, which in turn phosphorylates the Wiskott Aldrich Syndrome Protein (WASP. WASP is a key player in actin remodeling and cell migration. The reprogramed motility of infected macrophages observed in vitro correlates in vivo with enhanced macrophage infiltration in experimental tumors in Nef-transgenic mice compared to control mice.In conclusion, HIV infection of host target cells modifies their migration capacity; we infer that HIV-1 enhances virus spreading in confined environments by reducing T cells migration, and facilitates virus dissemination into different organs and tissues of the human body by enhancing macrophage mesenchymal migration.

  14. HIV-1 Nef control of cell signalling molecules: multiple strategies to promote virus replication

    Indian Academy of Sciences (India)

    Alison L Greenway; Gavan Holloway; Dale A McPhee; Phoebe Ellis; Alyssa Cornall; Michael Lidman

    2003-04-01

    HIV-1 has at its disposal numerous proteins encoded by its genome which provide the required arsenal to establish and maintain infection in its host for a considerable number of years. One of the most important and enigmatic of these proteins is Nef. The Nef protein of HIV-1 plays a fundamental role in the virus life cycle. This small protein of approximately 27 kDa is required for maximal virus replication and disease progression. The mechanisms by which it is able to act as a positive factor during virus replication is an area of intense research and although some controversy surrounds Nef much has been gauged as to how it functions. Its ability to modulate the expression of key cellular receptors important for cell activation and control signal transduction elements and events by interacting with numerous cellular kinases and signalling molecules, including members of the Src family kinases, leading to an effect on host cell function is likely to explain at least in part its role during infection and represents a finely tuned mechanism where this protein assists HIV-1 to control its host.

  15. CD4 and MHC class I down-modulation activities of nef alleles from brain- and lymphoid tissue-derived primary HIV-1 isolates

    Science.gov (United States)

    Gray, Lachlan R.; Gabuzda, Dana; Cowley, Daniel; Ellett, Anne; Chiavaroli, Lisa; Wesselingh, Steven L.; Churchill, Melissa J.; Gorry, Paul R.

    2015-01-01

    HIV-1 nef undergoes adaptive evolution in the CNS, reflecting altered requirements for HIV-1 replication in macrophages/microglia and brain-specific immune selection pressures. The role of Nef in HIV-1 neurotropism and the pathogenesis of HIV-associated dementia (HAD) is unclear. In this study, we characterized 82 nef alleles cloned from brain, CSF, spinal cord and blood/lymphoid tissue-derived HIV-1 isolates from 7 subjects with HAD. CNS isolate-derived nef alleles were genetically compartmentalized and had reduced sequence diversity compared to those from lymphoid tissue isolates. Defective nef alleles predominated in a brain-derived isolate from one of the 7 subjects (MACS2-br). The ability of Nef to down-modulate CD4 and MHC class 1 (MHC-1) was generally conserved among nef alleles from both CNS and lymphoid tissues. However, the potency of CD4 and MHC-1 down-modulation was variable, which was associated with sequence alterations known to influence these Nef functions. These results suggest that CD4 and MHC-1 down-modulation are highly conserved functions among nef alleles from CNS- and lymphoid tissue-derived HIV-1 isolates that may contribute to viral replication and escape from immune surveillance in the CNS. PMID:21165790

  16. In this issue: Nef, a lingering problem; are there better immunoglobulins for human use; and Takayasu arteritis, still a mystery.

    Science.gov (United States)

    Bot, Adrian

    2012-12-01

    The current issue of the International Reviews of Immunology brings the latest in three different areas of basic and clinical immunology. First, there is the lingering question of residual HIV-related pathology in patients on chronic antiretroviral therapy. The specific role of Nef in the macrophage-mediated disease manifested through lymphoma, metabolic disease and neurological disorder, is extensively discussed. A second topic is a clinical immunology one, a critical perspective on the efficacy and safety profile of various preparations of immunoglobulin products currently prescribed for a range of immunodeficiencies. Surprisingly, the authors showed that while the available preparations are equivalent from efficacy standpoint, they differ from the standpoint of toxicity. The third subject is again, in the arena of clinical immunology and deals with a relatively rare yet extremely puzzling disease -Takayasu arteritis -with a pathogenesis that needs elucidation and a dire need for better treatments. The authors provide a state of the art in terms of genetic association, other factors possibly involved in the onset and progression of this unusual inflammatory disease of large arteries, and provide a perspective on current standard of care and potential usefulness of TNF-α blockade as therapy for Takayasu arteritis.

  17. Rectal HSV-2 Infection May Increase Rectal SIV Acquisition Even in the Context of SIVΔnef Vaccination.

    Science.gov (United States)

    Guerra-Pérez, Natalia; Aravantinou, Meropi; Veglia, Filippo; Goode, Diana; Truong, Rosaline; Derby, Nina; Blanchard, James; Grasperge, Brooke; Gettie, Agegnehu; Robbiani, Melissa; Martinelli, Elena

    2016-01-01

    Prevalent HSV-2 infection increases the risk of HIV acquisition both in men and women even in asymptomatic subjects. Understanding the impact of HSV-2 on the mucosal microenvironment may help to identify determinants of susceptibility to HIV. Vaginal HSV-2 infection increases the frequency of cells highly susceptible to HIV in the vaginal tissue of women and macaques and this correlates with increased susceptibility to vaginal SHIV infection in macaques. However, the effect of rectal HSV-2 infection on HIV acquisition remains understudied. We developed a model of rectal HSV-2 infection in macaques in combination with rectal SIVmac239Δnef (SIVΔnef) vaccination and our results suggest that rectal HSV-2 infection may increase the susceptibility of macaques to rectal SIVmac239 wild-type (wt) infection even in SIVΔnef-infected animals. Rectal SIVΔnef infection/vaccination protected 7 out of 7 SIVΔnef-infected macaques from SIVmac239wt rectal infection (vs 12 out of 16 SIVΔnef-negative macaques), while 1 out of 3 animals co-infected with SIVΔnef and HSV-2 acquired SIVmac239wt infection. HSV-2/SIVmac239wt co-infected animals had increased concentrations of inflammatory factors in their plasma and rectal fluids and a tendency toward higher acute SIVmac239wt plasma viral load. However, they had higher blood CD4 counts and reduced depletion of CCR5+ CD4+ T cells compared to SIVmac239wt-only infected animals. Thus, rectal HSV-2 infection generates a pro-inflammatory environment that may increase susceptibility to rectal SIV infection and may impact immunological and virological parameters during acute SIV infection. Studies with larger number of animals are needed to confirm these findings.

  18. CD8 T Cell Response Maturation Defined by Anentropic Specificity and Repertoire Depth Correlates with SIVΔnef-induced Protection

    Science.gov (United States)

    Adnan, Sama; Colantonio, Arnaud D.; Yu, Yi; Gillis, Jacqueline; Wong, Fay E.; Becker, Ericka A.; Reeves, R. Keith; Lifson, Jeffrey D.; O’Connor, Shelby L.; Johnson, R. Paul

    2015-01-01

    The live attenuated simian immunodeficiency virus (LASIV) vaccine SIVΔnef is one of the most effective vaccines in inducing protection against wild-type lentiviral challenge, yet little is known about the mechanisms underlying its remarkable protective efficacy. Here, we exploit deep sequencing technology and comprehensive CD8 T cell epitope mapping to deconstruct the CD8 T cell response, to identify the regions of immune pressure and viral escape, and to delineate the effect of epitope escape on the evolution of the CD8 T cell response in SIVΔnef-vaccinated animals. We demonstrate that the initial CD8 T cell response in the acute phase of SIVΔnef infection is mounted predominantly against more variable epitopes, followed by widespread sequence evolution and viral escape. Furthermore, we show that epitope escape expands the CD8 T cell repertoire that targets highly conserved epitopes, defined as anentropic specificity, and generates de novo responses to the escaped epitope variants during the vaccination period. These results correlate SIVΔnef-induced protection with expanded anentropic specificity and increased response depth. Importantly, these findings render SIVΔnef, long the gold standard in HIV/SIV vaccine research, as a proof-of-concept vaccine that highlights the significance of the twin principles of anentropic specificity and repertoire depth in successful vaccine design. PMID:25688559

  19. CD8 T cell response maturation defined by anentropic specificity and repertoire depth correlates with SIVΔnef-induced protection.

    Directory of Open Access Journals (Sweden)

    Sama Adnan

    2015-02-01

    Full Text Available The live attenuated simian immunodeficiency virus (LASIV vaccine SIVΔnef is one of the most effective vaccines in inducing protection against wild-type lentiviral challenge, yet little is known about the mechanisms underlying its remarkable protective efficacy. Here, we exploit deep sequencing technology and comprehensive CD8 T cell epitope mapping to deconstruct the CD8 T cell response, to identify the regions of immune pressure and viral escape, and to delineate the effect of epitope escape on the evolution of the CD8 T cell response in SIVΔnef-vaccinated animals. We demonstrate that the initial CD8 T cell response in the acute phase of SIVΔnef infection is mounted predominantly against more variable epitopes, followed by widespread sequence evolution and viral escape. Furthermore, we show that epitope escape expands the CD8 T cell repertoire that targets highly conserved epitopes, defined as anentropic specificity, and generates de novo responses to the escaped epitope variants during the vaccination period. These results correlate SIVΔnef-induced protection with expanded anentropic specificity and increased response depth. Importantly, these findings render SIVΔnef, long the gold standard in HIV/SIV vaccine research, as a proof-of-concept vaccine that highlights the significance of the twin principles of anentropic specificity and repertoire depth in successful vaccine design.

  20. Sequence- and interactome-based prediction of viral protein hotspots targeting host proteins: a case study for HIV Nef.

    Directory of Open Access Journals (Sweden)

    Mahdi Sarmady

    Full Text Available Virus proteins alter protein pathways of the host toward the synthesis of viral particles by breaking and making edges via binding to host proteins. In this study, we developed a computational approach to predict viral sequence hotspots for binding to host proteins based on sequences of viral and host proteins and literature-curated virus-host protein interactome data. We use a motif discovery algorithm repeatedly on collections of sequences of viral proteins and immediate binding partners of their host targets and choose only those motifs that are conserved on viral sequences and highly statistically enriched among binding partners of virus protein targeted host proteins. Our results match experimental data on binding sites of Nef to host proteins such as MAPK1, VAV1, LCK, HCK, HLA-A, CD4, FYN, and GNB2L1 with high statistical significance but is a poor predictor of Nef binding sites on highly flexible, hoop-like regions. Predicted hotspots recapture CD8 cell epitopes of HIV Nef highlighting their importance in modulating virus-host interactions. Host proteins potentially targeted or outcompeted by Nef appear crowding the T cell receptor, natural killer cell mediated cytotoxicity, and neurotrophin signaling pathways. Scanning of HIV Nef motifs on multiple alignments of hepatitis C protein NS5A produces results consistent with literature, indicating the potential value of the hotspot discovery in advancing our understanding of virus-host crosstalk.

  1. Quantum criticality at the superconductor-insulator transition revealed by specific heat measurements.

    Science.gov (United States)

    Poran, S; Nguyen-Duc, T; Auerbach, A; Dupuis, N; Frydman, A; Bourgeois, Olivier

    2017-02-22

    The superconductor-insulator transition (SIT) is considered an excellent example of a quantum phase transition that is driven by quantum fluctuations at zero temperature. The quantum critical point is characterized by a diverging correlation length and a vanishing energy scale. Low-energy fluctuations near quantum criticality may be experimentally detected by specific heat, cp, measurements. Here we use a unique highly sensitive experiment to measure cp of two-dimensional granular Pb films through the SIT. The specific heat shows the usual jump at the mean field superconducting transition temperature marking the onset of Cooper pairs formation. As the film thickness is tuned towards the SIT, is relatively unchanged, while the magnitude of the jump and low-temperature specific heat increase significantly. This behaviour is taken as the thermodynamic fingerprint of quantum criticality in the vicinity of a quantum phase transition.

  2. Quantum criticality at the superconductor-insulator transition revealed by specific heat measurements

    Science.gov (United States)

    Poran, S.; Nguyen-Duc, T.; Auerbach, A.; Dupuis, N.; Frydman, A.; Bourgeois, Olivier

    2017-01-01

    The superconductor–insulator transition (SIT) is considered an excellent example of a quantum phase transition that is driven by quantum fluctuations at zero temperature. The quantum critical point is characterized by a diverging correlation length and a vanishing energy scale. Low-energy fluctuations near quantum criticality may be experimentally detected by specific heat, cp, measurements. Here we use a unique highly sensitive experiment to measure cp of two-dimensional granular Pb films through the SIT. The specific heat shows the usual jump at the mean field superconducting transition temperature marking the onset of Cooper pairs formation. As the film thickness is tuned towards the SIT, is relatively unchanged, while the magnitude of the jump and low-temperature specific heat increase significantly. This behaviour is taken as the thermodynamic fingerprint of quantum criticality in the vicinity of a quantum phase transition. PMID:28224994

  3. Critical Pedagogy, Internationalisation, and a Third Space: Cultural Tensions Revealed in Students' Discourse

    Science.gov (United States)

    Pitts, Margaret Jane; Brooks, Catherine F.

    2017-01-01

    Set within the context of a global pursuit towards the internationalisation of higher education, this paper critically examines student discourse in a globally connected classroom between learners in the USA and Singapore. It makes salient some of the cultural assumptions and tensions that undergird students' discourse in collaborative…

  4. Nef does not contribute to replication differences between R5 pre-AIDS and AIDS HIV-1 clones from patient ACH142

    Directory of Open Access Journals (Sweden)

    Rekosh David

    2008-05-01

    Full Text Available Abstract AIDS-associated, CCR5-tropic (R5 HIV-1 clones, isolated from a patient that never developed CXCR4-tropic HIV-1, replicate to a greater extent and cause greater cytopathic effects than R5 HIV-1 clones isolated before the onset of AIDS. Previously, we showed that HIV-1 Env substantially contributed to the enhanced replication of an AIDS clone. In order to determine if Nef makes a similar contribution, we cloned and phenotypically analyzed nef genes from a series of patient ACH142 derived R5 HIV-1 clones. The AIDS-associated Nef contains a series of residues found in Nef proteins from progressors 1. In contrast to other reports 123, this AIDS-associated Nef downmodulated MHC-I to a greater extent and CD4 less than pre-AIDS Nef proteins. Additionally, all Nef proteins enhanced infectivity similarly in a single round of replication. Combined with our previous study, these data show that evolution of the HIV-1 env gene, but not the nef gene, within patient ACH142 significantly contributed to the enhanced replication and cytopathic effects of the AIDS-associated R5 HIV-1 clone.

  5. Nef does not contribute to replication differences between R5 pre-AIDS and AIDS HIV-1 clones from patient ACH142

    Science.gov (United States)

    Olivieri, Kevin C; Scoggins, Robert M; Broderick, Brooks; Powell, Maria LC; Alexander, Melissa A; Hammarskjöld, Marie-Louise; Rekosh, David; Camerini, David

    2008-01-01

    AIDS-associated, CCR5-tropic (R5) HIV-1 clones, isolated from a patient that never developed CXCR4-tropic HIV-1, replicate to a greater extent and cause greater cytopathic effects than R5 HIV-1 clones isolated before the onset of AIDS. Previously, we showed that HIV-1 Env substantially contributed to the enhanced replication of an AIDS clone. In order to determine if Nef makes a similar contribution, we cloned and phenotypically analyzed nef genes from a series of patient ACH142 derived R5 HIV-1 clones. The AIDS-associated Nef contains a series of residues found in Nef proteins from progressors [1]. In contrast to other reports [1-3], this AIDS-associated Nef downmodulated MHC-I to a greater extent and CD4 less than pre-AIDS Nef proteins. Additionally, all Nef proteins enhanced infectivity similarly in a single round of replication. Combined with our previous study, these data show that evolution of the HIV-1 env gene, but not the nef gene, within patient ACH142 significantly contributed to the enhanced replication and cytopathic effects of the AIDS-associated R5 HIV-1 clone. PMID:18510766

  6. Mass balance of Nef glacier (Patagonia) by means of laser altimetry

    Science.gov (United States)

    Lopez, P.; Casassa, G.; Wendt, J.; Wendt, A.; Delclaux, F.

    2009-04-01

    Glaciers of Southern Patagonia Icefield (SPI) and Northern Patagonia Icefield (NPI) have shown an enhanced wasting and an increased melting in recent decades, mainly in reaction to regional warming. In consequence, water resources originating from those glaciers are also affected by their negative mass balance. Mass balance measurements provide information about the mass changes of both the accumulation and the ablation zones. A main limitation in attempting estimations of glacier mass balance of the NPI (4,197 km²) and the SPI (13,000 km²) is the difficulty in performing field observations, particularly within the accumulation areas, largely because of unfavourable meteorological conditions as well as the limitations due to the large size of the icefields. There is thus a need for carrying out detailed analyses of individual representative glaciers in Patagonia, covering both the ablation and accumulation areas. The Nef Glacier (138 km² in February 2005), is one of the largest and most representative glaciers of the eastern side of the NPI. During the last century it has been retreating and losing mass, and its evolution has been similar to other large glaciers of the NPI. Moreover, the Nef River is one of the most important tributaries of the Baker River, the largest drainage basin in the region and the river with the highest discharge in Chile. In this paper we present results of mass balance (2008 - 2009) of Nef glacier estimated using the geodetic method, where Digital Elevation Models (DEMs) of the glacier constructed at different dates are compared. The DEMs have been constructed using data from airborne laser altimetry with CECS Airborne Laser Scanner (Wendt et al., 2008), which has the advantage over airborne photogrammetry that it involves less data processing and practically no ground control, yielding excellent sub-meter precision.

  7. HIV-1 Group P is unable to antagonize human tetherin by Vpu, Env or Nef

    Directory of Open Access Journals (Sweden)

    Sauter Daniel

    2011-12-01

    Full Text Available Abstract Background A new subgroup of HIV-1, designated Group P, was recently detected in two unrelated patients of Cameroonian origin. HIV-1 Group P phylogenetically clusters with SIVgor suggesting that it is the result of a cross-species transmission from gorillas. Until today, HIV-1 Group P has only been detected in two patients, and its degree of adaptation to the human host is largely unknown. Previous data have shown that pandemic HIV-1 Group M, but not non-pandemic Group O or rare Group N viruses, efficiently antagonize the human orthologue of the restriction factor tetherin (BST-2, HM1.24, CD317 suggesting that primate lentiviruses may have to gain anti-tetherin activity for efficient spread in the human population. Thus far, three SIV/HIV gene products (vpu, nef and env are known to have the potential to counteract primate tetherin proteins, often in a species-specific manner. Here, we examined how long Group P may have been circulating in humans and determined its capability to antagonize human tetherin as an indicator of adaptation to humans. Results Our data suggest that HIV-1 Group P entered the human population between 1845 and 1989. Vpu, Env and Nef proteins from both Group P viruses failed to counteract human or gorilla tetherin to promote efficient release of HIV-1 virions, although both Group P Nef proteins moderately downmodulated gorilla tetherin from the cell surface. Notably, Vpu, Env and Nef alleles from the two HIV-1 P strains were all able to reduce CD4 cell surface expression. Conclusions Our analyses of the two reported HIV-1 Group P viruses suggest that zoonosis occurred in the last 170 years and further support that pandemic HIV-1 Group M strains are better adapted to humans than non-pandemic or rare Group O, N and P viruses. The inability to antagonize human tetherin may potentially explain the limited spread of HIV-1 Group P in the human population.

  8. Simulation of postsynaptic glutamate receptors reveals critical features of glutamatergic transmission.

    Directory of Open Access Journals (Sweden)

    Renaud Greget

    Full Text Available Activation of several subtypes of glutamate receptors contributes to changes in postsynaptic calcium concentration at hippocampal synapses, resulting in various types of changes in synaptic strength. Thus, while activation of NMDA receptors has been shown to be critical for long-term potentiation (LTP and long term depression (LTD of synaptic transmission, activation of metabotropic glutamate receptors (mGluRs has been linked to either LTP or LTD. While it is generally admitted that dynamic changes in postsynaptic calcium concentration represent the critical elements to determine the direction and amplitude of the changes in synaptic strength, it has been difficult to quantitatively estimate the relative contribution of the different types of glutamate receptors to these changes under different experimental conditions. Here we present a detailed model of a postsynaptic glutamatergic synapse that incorporates ionotropic and mGluR type I receptors, and we use this model to determine the role of the different receptors to the dynamics of postsynaptic calcium with different patterns of presynaptic activation. Our modeling framework includes glutamate vesicular release and diffusion in the cleft and a glutamate transporter that modulates extracellular glutamate concentration. Our results indicate that the contribution of mGluRs to changes in postsynaptic calcium concentration is minimal under basal stimulation conditions and becomes apparent only at high frequency of stimulation. Furthermore, the location of mGluRs in the postsynaptic membrane is also a critical factor, as activation of distant receptors contributes significantly less to calcium dynamics than more centrally located ones. These results confirm the important role of glutamate transporters and of the localization of mGluRs in postsynaptic sites in their signaling properties, and further strengthen the notion that mGluR activation significantly contributes to postsynaptic calcium

  9. Action potential processing in a detailed Purkinje cell model reveals a critical role for axonal compartmentalization

    Directory of Open Access Journals (Sweden)

    Stefano eMasoli

    2015-02-01

    Full Text Available The Purkinje cell (PC is among the most complex neurons in the brain and plays a critical role for cerebellar functioning. PCs operate as fast pacemakers modulated by synaptic inputs but can switch from simple spikes to complex bursts and, in some conditions, show bistability. In contrast to original works emphasizing dendritic Ca-dependent mechanisms, recent experiments have supported a primary role for axonal Na-dependent processing, which could effectively regulate spike generation and transmission to deep cerebellar nuclei (DCN. In order to account for the numerous ionic mechanisms involved (at present including Nav1.6, Cav2.1, Cav3.1, Cav3.2, Cav3.3, Kv1.1, Kv1.5, Kv3.3, Kv3.4, Kv4.3, KCa1.1, KCa2.2, KCa3.1, Kir2.x, HCN1, we have elaborated a multicompartmental model incorporating available knowledge on localization and gating of PC ionic channels. The axon, including initial segment (AIS and Ranvier nodes (RNs, proved critical to obtain appropriate pacemaking and firing frequency modulation. Simple spikes initiated in the AIS and protracted discharges were stabilized in the soma through Na-dependent mechanisms, while somato-dendritic Ca channels contributed to sustain pacemaking and to generate complex bursting at high discharge regimes. Bistability occurred only following Na and Ca channel down-regulation. In addition, specific properties in RNs K currents were required to limit spike transmission frequency along the axon. The model showed how organized electroresponsive functions could emerge from the molecular complexity of PCs and showed that the axon is fundamental to complement ionic channel compartmentalization enabling action potential processing and transmission of specific spike patterns to DCN.

  10. Pathogenicity and immunogenicity of attenuated, nef-deleted HIV-1 strains in vivo

    Directory of Open Access Journals (Sweden)

    Wesselingh Steven L

    2007-09-01

    Full Text Available Abstract In efforts to develop an effective vaccine, sterilizing immunity to primate lentiviruses has only been achieved by the use of live attenuated viruses carrying major deletions in nef and other accessory genes. Although live attenuated HIV vaccines are unlikely to be developed due to a myriad of safety concerns, opportunities exist to better understand the correlates of immune protection against HIV infection by studying rare cohorts of long-term survivors infected with attenuated, nef-deleted HIV strains such as the Sydney blood bank cohort (SBBC. Here, we review studies of viral evolution, pathogenicity, and immune responses to HIV infection in SBBC members. The studies show that potent, broadly neutralizing anti-HIV antibodies and robust CD8+ T-cell responses to HIV infection were not necessary for long-term control of HIV infection in a subset of SBBC members, and were not sufficient to prevent HIV sequence evolution, augmentation of pathogenicity and eventual progression of HIV infection in another subset. However, a persistent T-helper proliferative response to HIV p24 antigen was associated with long-term control of infection. Together, these results underscore the importance of the host in the eventual outcome of infection. Thus, whilst generating an effective antibody and CD8+ T-cell response are an essential component of vaccines aimed at preventing primary HIV infection, T-helper responses may be important in the generation of an effective therapeutic vaccine aimed at blunting chronic HIV infection.

  11. Molecular characterization of HIV-1 Nef and ACOT8 interaction: insights from in silico structural predictions and in vitro functional assays

    Science.gov (United States)

    Serena, Michela; Giorgetti, Alejandro; Busato, Mirko; Gasparini, Francesca; Diani, Erica; Romanelli, Maria Grazia; Zipeto, Donato

    2016-03-01

    HIV-1 Nef interacts with several cellular proteins, among which the human peroxisomal thioesterase 8 (ACOT8). This interaction may be involved in the endocytosis regulation of membrane proteins and might modulate lipid composition in membrane rafts. Nef regions involved in the interaction have been experimentally characterized, whereas structural details of the ACOT8 protein are unknown. The lack of structural information hampers the comprehension of the functional consequences of the complex formation during HIV-1 infection. We modelled, through in silico predictions, the ACOT8 structure and we observed a high charge complementarity between Nef and ACOT8 surfaces, which allowed the identification of the ACOT8 putative contact points involved in the interaction. The predictions were validated by in vitro assays through the development of ACOT8 deletion mutants. Coimmunoprecipitation and immunofluorescence analyses showed that ACOT8 Arg45-Phe55 and Arg86-Pro93 regions are involved in Nef association. In addition, K91S mutation abrogated the interaction with Nef, indicating that Lys91 plays a key role in the interaction. Finally, when associated with ACOT8, Nef may be preserved from degradation. These findings improve the comprehension of the association between HIV-1 Nef and ACOT8, helping elucidating the biological effect of their interaction.

  12. HIV-1 Nef enhances dendritic cell-mediated viral transmission to CD4+ T cells and promotes T-cell activation.

    Directory of Open Access Journals (Sweden)

    Corine St Gelais

    Full Text Available HIV-1 Nef enhances dendritic cell (DC-mediated viral transmission to CD4(+ T cells, but the underlying mechanism is not fully understood. It is also unknown whether HIV-1 infected DCs play a role in activating CD4(+ T cells and enhancing DC-mediated viral transmission. Here we investigated the role of HIV-1 Nef in DC-mediated viral transmission and HIV-1 infection of primary CD4(+ T cells using wild-type HIV-1 and Nef-mutated viruses. We show that HIV-1 Nef facilitated DC-mediated viral transmission to activated CD4(+ T cells. HIV-1 expressing wild-type Nef enhanced the activation and proliferation of primary resting CD4(+ T cells. However, when co-cultured with HIV-1-infected autologous DCs, there was no significant trend for infection- or Nef-dependent proliferation of resting CD4(+ T cells. Our results suggest an important role of Nef in DC-mediated transmission of HIV-1 to activated CD4(+ T cells and in the activation and proliferation of resting CD4(+ T cells, which likely contribute to viral pathogenesis.

  13. HIV-1 Infection of T Cells and Macrophages Are Differentially Modulated by Virion-Associated Hck: A Nef-Dependent Phenomenon

    Directory of Open Access Journals (Sweden)

    Gilda Tachedjian

    2013-09-01

    Full Text Available The proline repeat motif (PxxP of Nef is required for interaction with the SH3 domains of macrophage-specific Src kinase Hck. However, the implication of this interaction for viral replication and infectivity in macrophages and T lymphocytes remains unclear. Experiments in HIV-1 infected macrophages confirmed the presence of a Nef:Hck complex which was dependent on the Nef proline repeat motif. The proline repeat motif of Nef also enhanced both HIV-1 infection and replication in macrophages, and was required for incorporation of Hck into viral particles. Unexpectedly, wild-type Hck inhibited infection of macrophages, but Hck was shown to enhance infection of primary T lymphocytes. These results indicate that the interaction between Nef and Hck is important for Nef-dependent modulation of viral infectivity. Hck-dependent enhancement of HIV-1 infection of T cells suggests that Nef-Hck interaction may contribute to the spread of HIV-1 infection from macrophages to T cells by modulating events in the producer cell, virion and target cell.

  14. Detection of two amino acid deletions in HIV-1 Nef protein from Chinese former paid blood donors

    Institute of Scientific and Technical Information of China (English)

    SONG Yan-hui; WEI Min; MA Peng-fei; XING Hui; SI Xue-feng; TANG Hai-li; LI Hui-guang; SHAO Yi-ming

    2007-01-01

    @@ HIV-1 Nef protein plays an important role in HIV-1 pathogenesis, including downregulation of the cell surface CD4 and class Ⅰ major histocompatibility complex (MHC), enhancement of virion infectivity and alteration of a variety of cellular signal transduction pathways.

  15. Mutational analysis of Trypanosoma brucei RNA editing ligase reveals regions critical for interaction with KREPA2.

    Directory of Open Access Journals (Sweden)

    Vaibhav Mehta

    Full Text Available The Trypanosoma brucei parasite causes the vector-borne disease African sleeping sickness. Mitochondrial mRNAs of T. brucei undergo posttranscriptional RNA editing to make mature, functional mRNAs. The final step of this process is catalyzed by the essential ligase, T. brucei RNA Editing Ligase 1 (TbREL1 and the closely related T. brucei RNA Editing Ligase 2 (TbREL2. While other ligases such as T7 DNA ligase have both a catalytic and an oligonucleotide/oligosaccharide-binding (OB-fold domain, T. brucei RNA editing ligases contain only the catalytic domain. The OB-fold domain, which is required for interaction with the substrate RNA, is provided in trans by KREPA2 (for TbREL1 and KREPA1 (for TbREL2. KREPA2 enhancement of TbREL1 ligase activity is presumed to occur via an OB-fold-mediated increase in substrate specificity and catalysis. We characterized the interaction between TbREL1 and KREPA2 in vitro using full-length, truncated, and point-mutated ligases. As previously shown, our data indicate strong, specific stimulation of TbREL1 catalytic activity by KREPA2. We narrowed the region of contact to the final 59 C-terminal residues of TbREL1. Specifically, the TbREL1 C-terminal KWKE (441-444 sequence appear to coordinate the KREPA2-mediated enhancement of TbREL1 activities. N-terminal residues F206, T264 and Y275 are crucial for the overall activity of TbREL1, particularly for F206, a mutation of this residue also disrupts KREPA2 interaction. Thus, we have identified the critical TbREL1 regions and amino acids that mediate the KREPA2 interaction.

  16. Immunization of mice with the nef gene from Human Immunodeficiency Virus type 1: Study of immunological memory and long-term toxicology

    Directory of Open Access Journals (Sweden)

    Engström Gunnel

    2007-07-01

    Full Text Available Abstract Background The human immunodeficiency virus type 1 (HIV-1 regulatory protein, Nef, is an attractive vaccine target because it is involved in viral pathogenesis, is expressed early in the viral life cycle and harbors many T and B cell epitopes. Several clinical trials include gene-based vaccines encoding this protein. However, Nef has been shown to transform certain cell types in vitro. Based on these findings we performed a long-term toxicity and immunogenicity study of Nef, encoded either by Modified Vaccinia virus Ankara or by plasmid DNA. BALB/c mice were primed twice with either DNA or MVA encoding Nef and received a homologous or heterologous boost ten months later. In the meantime, the Nef-specific immune responses were monitored and at the time of sacrifice an extensive toxicological evaluation was performed, where presence of tumors and other pathological changes were assessed. Results The toxicological evaluation showed that immunization with MVAnef is safe and does not cause cellular transformation or other toxicity in somatic organs. Both DNAnef and MVAnef immunized animals developed potent Nef-specific cellular responses that declined to undetectable levels over time, and could readily be boosted after almost one year. This is of particular interest since it shows that plasmid DNA vaccine can also be used as a potent late booster of primed immune responses. We observed qualitative differences between the T cell responses induced by the two different vectors: DNA-encoded nef induced long-lasting CD8+ T cell memory responses, whereas MVA-encoded nef induced CD4+ T cell memory responses. In terms of the humoral immune responses, we show that two injections of MVAnef induce significant anti-Nef titers, while repeated injections of DNAnef do not. A single boost with MVAnef could enhance the antibody response following DNAnef prime to the same level as that observed in animals immunized repeatedly with MVAnef. We also demonstrate

  17. The Cytoplasmic Domain of CD4 Is Sufficient for Its Down-Regulation from the Cell Surface by Human Immunodeficiency Virus Type 1 Nef

    OpenAIRE

    S. J. Anderson; Lenburg, M; Landau, N R; Garcia, J. V.

    1994-01-01

    Human immunodeficiency virus type 1 Nef down-regulates surface expression of murine and human CD4 but not human CD8. We recently reported that the cytoplasmic domain of CD4 is required for its down-regulation by Nef. Using a chimeric molecule composed of the extracellular and transmembrane domains of human CD8 fused to the cytoplasmic domain of human CD4, we show here that the cytoplasmic domain of CD4 is sufficient for down-regulation by Nef. Since the cytoplasmic domain of CD4 is also the s...

  18. Mutagenesis of tGCN5 core region reveals two critical surface residues F90 and R140

    Energy Technology Data Exchange (ETDEWEB)

    Mehta, Kinjal Rajesh; Chan, Yan M.; Lee, Man X.; Yang, Ching Yao; Voloshchuk, Natalya [Department of Chemical and Biological Sciences, Polytechnic Institute of New York University, 6 MetroTech Center, Brooklyn, NY 11201 (United States); Montclare, Jin Kim, E-mail: jmontcla@poly.edu [Department of Chemical and Biological Sciences, Polytechnic Institute of New York University, 6 MetroTech Center, Brooklyn, NY 11201 (United States); Department of Biochemistry, SUNY-Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, NY 11203 (United States)

    2010-09-24

    Research highlights: {yields} Mutagenesis of the tGCN5 core region reveals two residues important for function. {yields} Developed a fluorescent lysate-based activity assay to assess mutants. {yields} Surface-exposed residues F90 and R140 of tGCN5 are critical for H3 acetylation. -- Abstract: Tetrahymena General Control Non-Derepressor 5 (tGCN5) is a critical regulator of gene transcription via acetylation of histones. Since the acetylation ability has been attributed to the 'core region', we perform mutagenesis of residues within the tGCN5 'core region' in order to identify those critical for function and stability. Residues that do not participate in catalysis are identified, mutated and characterized for activity, structure and thermodynamic stability. Variants I107V, Q114L, A121T and A130S maintain the acetylation function relative to wild-type tGCN5, while variants F90Y, F112R and R140H completely abolish function. Of the three non-functional variants, since F112 is mutated into a non-homologous charged residue, a loss in function is expected. However, the remaining two variants are mutated into homologous residues, suggesting that F90 and R140 are critical for the activity of tGCN5. While mutation to homologous residue maintains acetylation of histone H3 for the majority of the variants, the two surface-exposed residues, F90 and R140, appear to be essential for tGCN5 function, structure or stability.

  19. Characterization of CD8+ T cell differentiation following SIVΔnef vaccination by transcription factor expression profiling.

    Directory of Open Access Journals (Sweden)

    James M Billingsley

    2015-03-01

    Full Text Available The onset of protective immunity against pathogenic SIV challenge in SIVΔnef-vaccinated macaques is delayed for 15-20 weeks, a process that is related to qualitative changes in CD8+ T cell responses induced by SIVΔnef. As a novel approach to characterize cell differentiation following vaccination, we used multi-target qPCR to measure transcription factor expression in naïve and memory subsets of CD8++ T cells, and in SIV-specific CD8+ T cells obtained from SIVΔnef-vaccinated or wild type SIVmac239-infected macaques. Unsupervised clustering of expression profiles organized naïve and memory CD8+ T cells into groups concordant with cell surface phenotype. Transcription factor expression patterns in SIV-specific CD8+ T cells in SIVΔnef-vaccinated animals were distinct from those observed in purified CD8+ T cell subsets obtained from naïve animals, and were intermediate to expression profiles of purified central memory and effector memory T cells. Expression of transcription factors elicited by SIVΔnef vaccination also varied over time: cells obtained at later time points, temporally associated with greater protection, appeared more central-memory like than cells obtained at earlier time points, which appeared more effector memory-like. Expression of transcription factors associated with effector differentiation, such as ID2 and RUNX3, were decreased over time, while expression of transcription factors associated with quiescence or memory differentiation, such as TCF7, BCOR and EOMES, increased. CD8+ T cells specific for a more conserved epitope expressed higher levels of TBX21 and BATF, and appeared more effector-like than cells specific for an escaped epitope, consistent with continued activation by replicating vaccine virus. These data suggest transcription factor expression profiling is a novel method that can provide additional data complementary to the analysis of memory cell differentiation based on classical phenotypic markers

  20. Proteomic analysis reveals GIT1 as a novel mTOR complex component critical for mediating astrocyte survival.

    Science.gov (United States)

    Smithson, Laura J; Gutmann, David H

    2016-06-15

    As a critical regulator of cell growth, the mechanistic target of rapamycin (mTOR) protein operates as part of two molecularly and functionally distinct complexes. Herein, we demonstrate that mTOR complex molecular composition varies in different somatic tissues. In astrocytes and neural stem cells, we identified G-protein-coupled receptor kinase-interacting protein 1 (GIT1) as a novel mTOR-binding protein, creating a unique mTOR complex lacking Raptor and Rictor. Moreover, GIT1 binding to mTOR is regulated by AKT activation and is essential for mTOR-mediated astrocyte survival. Together, these data reveal that mTOR complex function is partly dictated by its molecuflar composition in different cell types.

  1. A whole-body model for glycogen regulation reveals a critical role for substrate cycling in maintaining blood glucose homeostasis.

    Directory of Open Access Journals (Sweden)

    Ke Xu

    2011-12-01

    Full Text Available Timely, and sometimes rapid, metabolic adaptation to changes in food supply is critical for survival as an organism moves from the fasted to the fed state, and vice versa. These transitions necessitate major metabolic changes to maintain energy homeostasis as the source of blood glucose moves away from ingested carbohydrates, through hepatic glycogen stores, towards gluconeogenesis. The integration of hepatic glycogen regulation with extra-hepatic energetics is a key aspect of these adaptive mechanisms. Here we use computational modeling to explore hepatic glycogen regulation under fed and fasting conditions in the context of a whole-body model. The model was validated against previous experimental results concerning glycogen phosphorylase a (active and glycogen synthase a dynamics. The model qualitatively reproduced physiological changes that occur during transition from the fed to the fasted state. Analysis of the model reveals a critical role for the inhibition of glycogen synthase phosphatase by glycogen phosphorylase a. This negative regulation leads to high levels of glycogen synthase activity during fasting conditions, which in turn increases substrate (futile cycling, priming the system for a rapid response once an external source of glucose is restored. This work demonstrates that a mechanistic understanding of the design principles used by metabolic control circuits to maintain homeostasis can benefit from the incorporation of mathematical descriptions of these networks into "whole-body" contextual models that mimic in vivo conditions.

  2. Hydrogen Exchange Mass Spectrometry of Related Proteins with Divergent Sequences: A Comparative Study of HIV-1 Nef Allelic Variants

    Science.gov (United States)

    Wales, Thomas E.; Poe, Jerrod A.; Emert-Sedlak, Lori; Morgan, Christopher R.; Smithgall, Thomas E.; Engen, John R.

    2016-06-01

    Hydrogen exchange mass spectrometry can be used to compare the conformation and dynamics of proteins that are similar in tertiary structure. If relative deuterium levels are measured, differences in sequence, deuterium forward- and back-exchange, peptide retention time, and protease digestion patterns all complicate the data analysis. We illustrate what can be learned from such data sets by analyzing five variants (Consensus G2E, SF2, NL4-3, ELI, and LTNP4) of the HIV-1 Nef protein, both alone and when bound to the human Hck SH3 domain. Regions with similar sequence could be compared between variants. Although much of the hydrogen exchange features were preserved across the five proteins, the kinetics of Nef binding to Hck SH3 were not the same. These observations may be related to biological function, particularly for ELI Nef where we also observed an impaired ability to downregulate CD4 surface presentation. The data illustrate some of the caveats that must be considered for comparison experiments and provide a framework for investigations of other protein relatives, families, and superfamilies with HX MS.

  3. Field transcriptome revealed critical developmental and physiological transitions involved in the expression of growth potential in japonica rice

    Directory of Open Access Journals (Sweden)

    Kamatsuki Kaori

    2011-01-01

    Full Text Available Abstract Background Plant growth depends on synergistic interactions between internal and external signals, and yield potential of crops is a manifestation of how these complex factors interact, particularly at critical stages of development. As an initial step towards developing a systems-level understanding of the biological processes underlying the expression of overall agronomic potential in cereal crops, a high-resolution transcriptome analysis of rice was conducted throughout life cycle of rice grown under natural field conditions. Results A wide range of gene expression profiles based on 48 organs and tissues at various developmental stages identified 731 organ/tissue specific genes as well as 215 growth stage-specific expressed genes universally in leaf blade, leaf sheath, and root. Continuous transcriptome profiling of leaf from transplanting until harvesting further elucidated the growth-stage specificity of gene expression and uncovered two major drastic changes in the leaf transcriptional program. The first major change occurred before the panicle differentiation, accompanied by the expression of RFT1, a putative florigen gene in long day conditions, and the downregulation of the precursors of two microRNAs. This transcriptome change was also associated with physiological alterations including phosphate-homeostasis state as evident from the behavior of several key regulators such as miR399. The second major transcriptome change occurred just after flowering, and based on analysis of sterile mutant lines, we further revealed that the formation of strong sink, i.e., a developing grain, is not the major cause but is rather a promoter of this change. Conclusions Our study provides not only the genetic basis for functional genomics in rice but also new insight into understanding the critical physiological processes involved in flowering and seed development, that could lead to novel strategies for optimizing crop productivity.

  4. HLA-associated immune escape pathways in HIV-1 subtype B Gag, Pol and Nef proteins.

    Directory of Open Access Journals (Sweden)

    Zabrina L Brumme

    Full Text Available BACKGROUND: Despite the extensive genetic diversity of HIV-1, viral evolution in response to immune selective pressures follows broadly predictable mutational patterns. Sites and pathways of Human Leukocyte-Antigen (HLA-associated polymorphisms in HIV-1 have been identified through the analysis of population-level data, but the full extent of immune escape pathways remains incompletely characterized. Here, in the largest analysis of HIV-1 subtype B sequences undertaken to date, we identify HLA-associated polymorphisms in the three HIV-1 proteins most commonly considered in cellular-based vaccine strategies. Results are organized into protein-wide escape maps illustrating the sites and pathways of HLA-driven viral evolution. METHODOLOGY/PRINCIPAL FINDINGS: HLA-associated polymorphisms were identified in HIV-1 Gag, Pol and Nef in a multicenter cohort of >1500 chronically subtype-B infected, treatment-naïve individuals from established cohorts in Canada, the USA and Western Australia. At q< or =0.05, 282 codons commonly mutating under HLA-associated immune pressures were identified in these three proteins. The greatest density of associations was observed in Nef (where close to 40% of codons exhibited a significant HLA association, followed by Gag then Pol (where approximately 15-20% of codons exhibited HLA associations, confirming the extensive impact of immune selection on HIV evolution and diversity. Analysis of HIV codon covariation patterns identified over 2000 codon-codon interactions at q< or =0.05, illustrating the dense and complex networks of linked escape and secondary/compensatory mutations. CONCLUSIONS/SIGNIFICANCE: The immune escape maps and associated data are intended to serve as a user-friendly guide to the locations of common escape mutations and covarying codons in HIV-1 subtype B, and as a resource facilitating the systematic identification and classification of immune escape mutations. These resources should facilitate

  5. Soil water stable isotopes reveal evaporation dynamics at the soil-plant-atmosphere interface of the critical zone

    Science.gov (United States)

    Sprenger, Matthias; Tetzlaff, Doerthe; Soulsby, Chris

    2017-07-01

    Understanding the influence of vegetation on water storage and flux in the upper soil is crucial in assessing the consequences of climate and land use change. We sampled the upper 20 cm of podzolic soils at 5 cm intervals in four sites differing in their vegetation (Scots Pine (Pinus sylvestris) and heather (Calluna sp. and Erica Sp)) and aspect. The sites were located within the Bruntland Burn long-term experimental catchment in the Scottish Highlands, a low energy, wet environment. Sampling took place on 11 occasions between September 2015 and September 2016 to capture seasonal variability in isotope dynamics. The pore waters of soil samples were analyzed for their isotopic composition (δ2H and δ18O) with the direct-equilibration method. Our results show that the soil waters in the top soil are, despite the low potential evaporation rates in such northern latitudes, kinetically fractionated compared to the precipitation input throughout the year. This fractionation signal decreases within the upper 15 cm resulting in the top 5 cm being isotopically differentiated to the soil at 15-20 cm soil depth. There are significant differences in the fractionation signal between soils beneath heather and soils beneath Scots pine, with the latter being more pronounced. But again, this difference diminishes within the upper 15 cm of soil. The enrichment in heavy isotopes in the topsoil follows a seasonal hysteresis pattern, indicating a lag time between the fractionation signal in the soil and the increase/decrease of soil evaporation in spring/autumn. Based on the kinetic enrichment of the soil water isotopes, we estimated the soil evaporation losses to be about 5 and 10 % of the infiltrating water for soils beneath heather and Scots pine, respectively. The high sampling frequency in time (monthly) and depth (5 cm intervals) revealed high temporal and spatial variability of the isotopic composition of soil waters, which can be critical, when using stable isotopes as tracers

  6. Inefficient Nef-mediated downmodulation of CD3 and MHC-I correlates with loss of CD4+T cells in natural SIV infection.

    Directory of Open Access Journals (Sweden)

    Michael Schindler

    2008-07-01

    Full Text Available Recent data suggest that Nef-mediated downmodulation of TCR-CD3 may protect SIVsmm-infected sooty mangabeys (SMs against the loss of CD4+ T cells. However, the mechanisms underlying this protective effect remain unclear. To further assess the role of Nef in nonpathogenic SIV infection, we cloned nef alleles from 11 SIVsmm-infected SMs with high (>500 and 15 animals with low (<500 CD4+ T-cells/microl in bulk into proviral HIV-1 IRES/eGFP constructs and analyzed their effects on the phenotype, activation, and apoptosis of primary T cells. We found that not only efficient Nef-mediated downmodulation of TCR-CD3 but also of MHC-I correlated with preserved CD4+ T cell counts, as well as with high numbers of Ki67+CD4+ and CD8+CD28+ T cells and reduced CD95 expression by CD4+ T cells. Moreover, effective MHC-I downregulation correlated with low proportions of effector and high percentages of naïve and memory CD8+ T cells. We found that T cells infected with viruses expressing Nef alleles from the CD4low SM group expressed significantly higher levels of the CD69, interleukin (IL-2 and programmed death (PD-1 receptors than those expressing Nefs from the CD4high group. SIVsmm Nef alleles that were less active in downmodulating TCR-CD3 were also less potent in suppressing the activation of virally infected T cells and subsequent cell death. However, only nef alleles from a single animal with very low CD4+ T cell counts rendered T cells hyper-responsive to activation, similar to those of HIV-1. Our data suggest that Nef may protect the natural hosts of SIV against the loss of CD4+ T cells by at least two mechanisms: (i downmodulation of TCR-CD3 to prevent activation-induced cell death and to suppress the induction of PD-1 that may impair T cell function and survival, and (ii downmodulation of MHC-I to reduce CTL lysis of virally infected CD4+ T cells and/or bystander CD8+ T cell activation.

  7. Formation of a unique cluster of G-quadruplex structures in the HIV-1 Nef coding region: implications for antiviral activity.

    Directory of Open Access Journals (Sweden)

    Rosalba Perrone

    Full Text Available G-quadruplexes are tetraplex structures of nucleic acids that can form in G-rich sequences. Their presence and functional role have been established in telomeres, oncogene promoters and coding regions of the human chromosome. In particular, they have been proposed to be directly involved in gene regulation at the level of transcription. Because the HIV-1 Nef protein is a fundamental factor for efficient viral replication, infectivity and pathogenesis in vitro and in vivo, we investigated G-quadruplex formation in the HIV-1 nef gene to assess the potential for viral inhibition through G-quadruplex stabilization. A comprehensive computational analysis of the nef coding region of available strains showed the presence of three conserved sequences that were uniquely clustered. Biophysical testing proved that G-quadruplex conformations were efficiently stabilized or induced by G-quadruplex ligands in all three sequences. Upon incubation with a G-quadruplex ligand, Nef expression was reduced in a reporter gene assay and Nef-dependent enhancement of HIV-1 infectivity was significantly repressed in an antiviral assay. These data constitute the first evidence of the possibility to regulate HIV-1 gene expression and infectivity through G-quadruplex targeting and therefore open a new avenue for viral treatment.

  8. RNA-Seq Analysis of Sulfur-Deprived Chlamydomonas Cells Reveals Aspects of Acclimation Critical for Cell Survival[W

    Science.gov (United States)

    González-Ballester, David; Casero, David; Cokus, Shawn; Pellegrini, Matteo; Merchant, Sabeeha S.; Grossman, Arthur R.

    2010-01-01

    The Chlamydomonas reinhardtii transcriptome was characterized from nutrient-replete and sulfur-depleted wild-type and snrk2.1 mutant cells. This mutant is null for the regulatory Ser-Thr kinase SNRK2.1, which is required for acclimation of the alga to sulfur deprivation. The transcriptome analyses used microarray hybridization and RNA-seq technology. Quantitative RT-PCR evaluation of the results obtained by these techniques showed that RNA-seq reports a larger dynamic range of expression levels than do microarray hybridizations. Transcripts responsive to sulfur deprivation included those encoding proteins involved in sulfur acquisition and assimilation, synthesis of sulfur-containing metabolites, Cys degradation, and sulfur recycling. Furthermore, we noted potential modifications of cellular structures during sulfur deprivation, including the cell wall and complexes associated with the photosynthetic apparatus. Moreover, the data suggest that sulfur-deprived cells accumulate proteins with fewer sulfur-containing amino acids. Most of the sulfur deprivation responses are controlled by the SNRK2.1 protein kinase. The snrk2.1 mutant exhibits a set of unique responses during both sulfur-replete and sulfur-depleted conditions that are not observed in wild-type cells; the inability of this mutant to acclimate to S deprivation probably leads to elevated levels of singlet oxygen and severe oxidative stress, which ultimately causes cell death. The transcriptome results for wild-type and mutant cells strongly suggest the occurrence of massive changes in cellular physiology and metabolism as cells become depleted for sulfur and reveal aspects of acclimation that are likely critical for cell survival. PMID:20587772

  9. Mutational analysis of an archaeal minichromosome maintenance protein exterior hairpin reveals critical residues for helicase activity and DNA binding

    Directory of Open Access Journals (Sweden)

    Brewster Aaron S

    2010-08-01

    Full Text Available Abstract Background The mini-chromosome maintenance protein (MCM complex is an essential replicative helicase for DNA replication in Archaea and Eukaryotes. While the eukaryotic complex consists of six homologous proteins (MCM2-7, the archaeon Sulfolobus solfataricus has only one MCM protein (ssoMCM, six subunits of which form a homohexamer. We have recently reported a 4.35Å crystal structure of the near full-length ssoMCM. The structure reveals a total of four β-hairpins per subunit, three of which are located within the main channel or side channels of the ssoMCM hexamer model generated based on the symmetry of the N-terminal Methanothermobacter thermautotrophicus (mtMCM structure. The fourth β-hairpin, however, is located on the exterior of the hexamer, near the exit of the putative side channels and next to the ATP binding pocket. Results In order to better understand this hairpin's role in DNA binding and helicase activity, we performed a detailed mutational and biochemical analysis of nine residues on this exterior β-hairpin (EXT-hp. We examined the activities of the mutants related to their helicase function, including hexamerization, ATPase, DNA binding and helicase activities. The assays showed that some of the residues on this EXT-hp play a role for DNA binding as well as for helicase activity. Conclusions These results implicate several current theories regarding helicase activity by this critical hexameric enzyme. As the data suggest that EXT-hp is involved in DNA binding, the results reported here imply that the EXT-hp located near the exterior exit of the side channels may play a role in contacting DNA substrate in a manner that affects DNA unwinding.

  10. Promoter analysis reveals critical roles for SMAD-3 and ATF-2 in expression of IL-23 p19 in macrophages.

    Science.gov (United States)

    Al-Salleeh, Fahd; Petro, Thomas M

    2008-10-01

    IL-23 p19/p40, produced by macrophages and dendritic cells, is critical for development of Th17 in several autoimmune diseases. In this study, bone marrow-derived (BMM) and splenic macrophages (SPM) from SJL/J mice, susceptible to autoimmune demyelinating disease following Theiler's virus (TMEV) infection, expressed IL-23 in response to TMEV. We identified potential binding sites for IFN response factor (IRF)-3 (nt -734 to -731), Sma- and Mad-related protein (SMAD)-3 (nt -584 to -581), activating transcription factor (ATF)-2 (nt -571 to -568), IRF-7 (nt -533 to-525), and NF-kappaB (nt -215 to -209) in the murine p19 promoter. The p19(prom) in the pGL3 promoter-reporter vector responded to TMEV or poly(I:C), a TLR3 agonist in the RAW264.7 macrophage cell line. Deletions upstream from the IRF-3 site and mutations at the IRF-3, SMAD-3, ATF-2, or NF-kappaB, but not the IRF-7, sites significantly reduced promoter activity. ATF-2 or SMAD-3, but not IRF-3, short-hairpin RNA reduced p19 promoter activity and protein expression in RAW264.7 cells responding to TMEV. Chromosomal DNA immunoprecipitation assays revealed that SMAD-3 and ATF-2 bind to the endogenous p19 promoter in RAW264.7 cells and SJL/J SPM following challenge with TMEV. TGF-beta1, which activates SMAD-3, was induced in RAW264.7 cells, BMM, and SPM by TMEV. Neutralizing Ab to TGF-beta1 eliminated TMEV-induced IL-23 production and SMAD-3 activation in RAW264.7 cells, BMM, and SPM. Activation of ATF-2 was JNK, but not p38 or ERK MAPK dependent. Inhibition of the JNK, but also the ERK MAPK pathways decreased expression of p19. These results suggest that ATF-2 and SMAD-3 are transcription factors, which are, in addition to NF-kappaB, essential for IL-23 p19 expression.

  11. Phenotype and envelope gene diversity of nef-deleted HIV-1 isolated from long-term survivors infected from a single source

    Directory of Open Access Journals (Sweden)

    Sullivan John S

    2007-07-01

    Full Text Available Abstract Background The Sydney blood bank cohort (SBBC of long-term survivors consists of multiple individuals infected with attenuated, nef-deleted variants of human immunodeficiency virus type 1 (HIV-1 acquired from a single source. Long-term prospective studies have demonstrated that the SBBC now comprises slow progressors (SP as well as long-term nonprogressors (LTNP. Convergent evolution of nef sequences in SBBC SP and LTNP indicates the in vivo pathogenicity of HIV-1 in SBBC members is dictated by factors other than nef. To better understand mechanisms underlying the pathogenicity of nef-deleted HIV-1, we examined the phenotype and env sequence diversity of sequentially isolated viruses (n = 2 from 3 SBBC members. Results The viruses characterized here were isolated from two SP spanning a three or six year period during progressive HIV-1 infection (subjects D36 and C98, respectively and from a LTNP spanning a two year period during asymptomatic, nonprogressive infection (subject C18. Both isolates from D36 were R5X4 phenotype and, compared to control HIV-1 strains, replicated to low levels in peripheral blood mononuclear cells (PBMC. In contrast, both isolates from C98 and C18 were CCR5-restricted. Both viruses isolated from C98 replicated to barely detectable levels in PBMC, whereas both viruses isolated from C18 replicated to low levels, similar to those isolated from D36. Analysis of env by V1V2 and V3 heteroduplex tracking assay, V1V2 length polymorphisms, sequencing and phylogenetic analysis showed distinct intra- and inter-patient env evolution. Conclusion Independent evolution of env despite convergent evolution of nef may contribute to the in vivo pathogenicity of nef-deleted HIV-1 in SBBC members, which may not necessarily be associated with changes in replication capacity or viral coreceptor specificity.

  12. Evolution of the HIV-1 nef gene in HLA-B*57 Positive Elite Suppressors

    OpenAIRE

    Siliciano Robert F; Ray Stuart C; Bailey Justin R; O'Connell Karen A; Brennan Timothy P; Salgado Maria; Blankson Joel N

    2010-01-01

    Abstract Elite controllers or suppressors (ES) are HIV-1 infected patients who maintain viral loads of < 50 copies/ml without antiretroviral therapy. CD8+ T cells are thought to play a key role in the control of viral replication and exert selective pressure on gag and nef in HLA-B*57 positive ES. We previously showed evolution in the gag gene of ES which surprisingly was mostly due to synonymous mutations rather than non-synonymous mutation in targeted CTL epitopes. This finding could be the...

  13. Overlapping cis sites used for splicing of HIV-1 env/nef and rev mRNAs.

    Science.gov (United States)

    Swanson, A K; Stoltzfus, C M

    1998-12-18

    Alternative splicing is used to generate more than 30 human immunodeficiency virus type 1 (HIV-1) spliced and unspliced mRNAs from a single primary transcript. The abundance of HIV-1 mRNAs is determined by the efficiencies with which its different 5' and 3' splice sites are used. Three splice sites (A4c, A4a, and A4b) are upstream of the rev initiator AUG. RNAs spliced at A4c, A4a, and A4b are used as mRNAs for Rev. Another 3' splice site (A5) is immediately downstream of the rev initiator. RNAs spliced at A5 are used as mRNAs for Env and Nef. In this report, primer extension analysis of splicing intermediates was used to show that there are eight branch points in this region, all of which map to adenosine residues. In addition, cis elements recognized by the cellular splicing machinery overlap; the two most 3' branch points overlap with the AG dinucleotides at rev 3' splice sites A4a and A4b. Competition of the overlapping cis sites for different splicing factors may play a role in maintaining the appropriate balance of mRNAs in HIV-1-infected cells. In support of this possibility, mutations at rev 3' splice site A4b AG dinucleotide dramatically increased splicing of the env/nef 3' splice site A5. This correlated with increased usage of the four most 3' branch points, which include those within the rev 3' splice site AG dinucleotides. Consistent with these results, analysis of a mutant in which three of the four env/nef branch points were inactivated indicated that use of splice site A5 was inhibited and splicing was shifted predominantly to the most 5' rev 3' splice site A4c with preferential use of the two most 5' branch points. Our results suggest that spliceosomes formed at rev A4a-4b, rev A4c, and env/nef A5 3' splice sites each recognize different subsets of the eight branch point sequences.

  14. High-level HIV-1 Nef transient expression in Nicotiana benthamiana using the P19 gene silencing suppressor protein of Artichoke Mottled Crinckle Virus

    Directory of Open Access Journals (Sweden)

    Bianco Linda

    2009-11-01

    Full Text Available Abstract Background In recent years, different HIV antigens have been successfully expressed in plants by either stable transformation or transient expression systems. Among HIV proteins, Nef is considered a promising target for the formulation of a multi-component vaccine due to its implication in the first steps of viral infection. Attempts to express Nef as a single protein product (not fused to a stabilizing protein in transgenic plants resulted in disappointingly low yields (about 0.5% of total soluble protein. In this work we describe a transient expression system based on co-agroinfiltration of plant virus gene silencing suppressor proteins in Nicotiana benthamiana, followed by a two-step affinity purification protocol of plant-derived Nef. Results The effect of three gene silencing viral suppressor proteins (P25 of Potato Virus X, P19 of either Artichoke Mottled Crinckle virus and Tomato Bushy Stunt virus on Nef transient expression yield was evaluated. The P19 protein of Artichoke Mottled Crinckle virus (AMCV-P19 gave the highest expression yield in vacuum co-agroinfiltration experiments reaching 1.3% of total soluble protein, a level almost three times higher than that previously reported in stable transgenic plants. The high yield observed in the co-agroinfiltrated plants was correlated to a remarkable decrease of Nef-specific small interfering RNAs (siRNAs indicating an effective modulation of RNA silencing mechanisms by AMCV-P19. Interestingly, we also showed that expression levels in top leaves of vacuum co-agroinfiltrated plants were noticeably reduced compared to bottom leaves. Moreover, purification of Nef from agroinfiltrated tissue was achieved by a two-step immobilized metal ion affinity chromatography protocol with yields of 250 ng/g of fresh tissue. Conclusion We demonstrated that expression level of HIV-1 Nef in plant can be improved using a transient expression system enhanced by the AMCV-P19 gene silencing suppressor

  15. Sequence heterogeneity in human immunodeficiency virus type 1 nef in patients presenting with rapid progression and delayed progression to AIDS.

    Science.gov (United States)

    Gupta, Poonam; Husain, Mohammad; Hans, Charoo; Ram, Hanu; Verma, Supriya Singh; Misbah, Mohammad; Chauhan, L S; Rai, Arvind

    2014-09-01

    Genetic heterogeneity in the nef genes from human immunodeficiency virus type 1 (HIV-1)-infected rapid progressors (RPs) and long-term nonprogressors (LTNPs) was analyzed to identify various amino acid substitutions responsible for the discernible difference in disease progression. It was found that the majority of the strains characterized belonged to subtype C, followed by several BC recombinants and subtype A1. Complete nef subtype C sequences from 33 RPs and seven LTNPs were compared, and it was observed that, in the majority of the sequences from both groups, highly conserved functional motifs showed subtle changes. However, drastic changes were observed in two isolates from LTNPs where the arginine cluster was deleted, while in one of them, additionally, acidic residues were replaced by basic residues (EEEEE→RK(R)KKE). The deletion of the arginine cluster and the mutation of acidic residues to basic residues are predicted to delay disease development by abolishing CD4 downmodulation and causing diminution of major histocompatibility complex class I (MHC-I) downregulation, respectively. Nonetheless, this is an exclusive finding in these LTNPs, which necessitates their analysis at the functional level. The synonymous-to-nonsynonymous substitution ratio was greater than one in both of the groups, suggesting amino acid sequence conservation and functional robustness. Interpatient nucleotide distance within the group and between the two groups showed very little variation, confirming genetic relatedness among isolates.

  16. The human immunodeficiency virus (HIV) Rev-binding protein (HRB) is a co-factor for HIV-1 Nef-mediated CD4 downregulation.

    Science.gov (United States)

    Landi, Alessia; Timermans, Cristina Garcia; Naessens, Evelien; Vanderstraeten, Hanne; Stove, Veronique; Verhasselt, Bruno

    2016-03-01

    Human immunodeficiency virus type 1 (HIV-1)-mediated CD4 downregulation is an important determinant of viral replication in vivo. Research on cellular co-factors involved in this process could lead to the identification of potential therapeutic targets. We found that CD4 surface levels were significantly higher in HIV-1-infected cells knocked-down for the HIV Rev-binding protein (HRB) compared with control cells. HRB knock-down affected CD4 downregulation induced by Nef but not by HIV-1 Vpu. Interestingly, the knock-down of the related protein HRBL (HRB-like), but not of the HRB interaction partner EPS15 (epidermal growth factor receptor pathway substrate 15), increased CD4 levels in Vpu-expressing cells significantly. Both of these proteins are known to be involved in HIV-1-mediated CD4 downregulation as co-factors of HIV-1 Nef. These results identify HRB as a previously unknown co-factor for HIV-1 Nef-mediated CD4 downregulation and highlight differences with the related protein HRBL, which affects the CD4 downregulation in a dual role as co-factor of both HIV-1 Nef and Vpu.

  17. Ear manipulations reveal a critical period for survival and dendritic development at the single-cell level in Mauthner neurons.

    Science.gov (United States)

    Elliott, Karen L; Houston, Douglas W; DeCook, Rhonda; Fritzsch, Bernd

    2015-12-01

    Second-order sensory neurons are dependent on afferents from the sense organs during a critical period in development for their survival and differentiation. Past research has mostly focused on whole populations of neurons, hampering progress in understanding the mechanisms underlying these critical phases. To move toward a better understanding of the molecular and cellular basis of afferent-dependent neuronal development, we developed a new model to study the effects of ear removal on a single identifiable cell in the hindbrain of a frog, the Mauthner cell. Ear extirpation at various stages of Xenopus laevis development defines a critical period of progressively-reduced dependency of Mauthner cell survival/differentiation on the ear afferents. Furthermore, ear removal results in a progressively decreased reduction in the number of dendritic branches. Conversely, addition of an ear results in an increase in the number of dendritic branches. These results suggest that the duration of innervation and the number of inner ear afferents play a quantitative role in Mauthner cell survival/differentiation, including dendritic development.

  18. Theory before practice: implicit assumptions about clinical nursing education in Australia as revealed through a shared critical reflection.

    Science.gov (United States)

    Grealish, Laurie; Smale, Lacey Anne

    2011-08-01

    The transfer of nursing education into the higher education sector occurred over a 10-year period in Australia (1985-1994). Australian nurse leaders settled on a single outcome measure to be applied for all nursing graduates in the form of national competency standards. While this move enabled diversity, the lack of consistency in curriculum design has subsequently led to increasing confusion for clinicians who support students' learning in clinical placements. Using a shared critical reflection method, the authors reviewed (1) the evaluation comments from nurses in one nursing unit of a hospital in one Australian jurisdiction and (2) an historical review of nursing literature at the time of the transfer of nursing education into the higher education sector. The reflection suggests that the aim of the transfer, to create critical thinking graduates, has been undermined by the implicit clinical education practices that have since emerged. In order to address the contemporary challenges for clinical staff working with students from multiple universities, as well as increased student numbers to address the nursing shortage, we recommend a new approach to curriculum design: a national clinical curriculum drawn from social, as well as cognitive, learning theory that at once informs clinicians of students' potential abilities and provides the scope to accommodate the increasingly difficult and critical learning requirements of tertiary-based nursing students.

  19. Persistent Low-Level Replication of SIVΔnef Drives Maturation of Antibody and CD8 T Cell Responses to Induce Protective Immunity against Vaginal SIV Infection.

    Directory of Open Access Journals (Sweden)

    Sama Adnan

    2016-12-01

    Full Text Available Defining the correlates of immune protection conferred by SIVΔnef, the most effective vaccine against SIV challenge, could enable the design of a protective vaccine against HIV infection. Here we provide a comprehensive assessment of immune responses that protect against SIV infection through detailed analyses of cellular and humoral immune responses in the blood and tissues of rhesus macaques vaccinated with SIVΔnef and then vaginally challenged with wild-type SIV. Despite the presence of robust cellular immune responses, animals at 5 weeks after vaccination displayed only transient viral suppression of challenge virus, whereas all macaques challenged at weeks 20 and 40 post-SIVΔnef vaccination were protected, as defined by either apparent sterile protection or significant suppression of viremia in infected animals. Multiple parameters of CD8 T cell function temporally correlated with maturation of protection, including polyfunctionality, phenotypic differentiation, and redistribution to gut and lymphoid tissues. Importantly, we also demonstrate the induction of a tissue-resident memory population of SIV-specific CD8 T cells in the vaginal mucosa, which was dependent on ongoing low-level antigenic stimulation. Moreover, we show that vaginal and serum antibody titers inversely correlated with post-challenge peak viral load, and we correlate the accumulation and affinity maturation of the antibody response to the duration of the vaccination period as well as to the SIVΔnef antigenic load. In conclusion, maturation of SIVΔnef-induced CD8 T cell and antibody responses, both propelled by viral persistence in the gut mucosa and secondary lymphoid tissues, results in protective immune responses that are able to interrupt viral transmission at mucosal portals of entry as well as potential sites of viral dissemination.

  20. Emergent Self-Organized Criticality in Gene Expression Dynamics: Temporal Development of Global Phase Transition Revealed in a Cancer Cell Line

    Science.gov (United States)

    Tsuchiya, Masa; Giuliani, Alessandro; Hashimoto, Midori; Erenpreisa, Jekaterina; Yoshikawa, Kenichi

    2015-01-01

    Background The underlying mechanism of dynamic control of the genome-wide expression is a fundamental issue in bioscience. We addressed it in terms of phase transition by a systemic approach based on both density analysis and characteristics of temporal fluctuation for the time-course mRNA expression in differentiating MCF-7 breast cancer cells. Methodology In a recent work, we suggested criticality as an essential aspect of dynamic control of genome-wide gene expression. Criticality was evident by a unimodal-bimodal transition through flattened unimodal expression profile. The flatness on the transition suggests the existence of a critical transition at which up- and down-regulated expression is balanced. Mean field (averaging) behavior of mRNAs based on the temporal expression changes reveals a sandpile type of transition in the flattened profile. Furthermore, around the transition, a self-similar unimodal-bimodal transition of the whole expression occurs in the density profile of an ensemble of mRNA expression. These singular and scaling behaviors identify the transition as the expression phase transition driven by self-organized criticality (SOC). Principal Findings Emergent properties of SOC through a mean field approach are revealed: i) SOC, as a form of genomic phase transition, consolidates distinct critical states of expression, ii) Coupling of coherent stochastic oscillations between critical states on different time-scales gives rise to SOC, and iii) Specific gene clusters (barcode genes) ranging in size from kbp to Mbp reveal similar SOC to genome-wide mRNA expression and ON-OFF synchronization to critical states. This suggests that the cooperative gene regulation of topological genome sub-units is mediated by the coherent phase transitions of megadomain-scaled conformations between compact and swollen chromatin states. Conclusion and Significance In summary, our study provides not only a systemic method to demonstrate SOC in whole-genome expression

  1. Atomic structure of recombinant thaumatin II reveals flexible conformations in two residues critical for sweetness and three consecutive glycine residues.

    Science.gov (United States)

    Masuda, Tetsuya; Mikami, Bunzo; Tani, Fumito

    2014-11-01

    Thaumatin, an intensely sweet-tasting protein used as a sweetener, elicits a sweet taste at 50 nM. Although two major variants designated thaumatin I and thaumatin II exist in plants, there have been few dedicated thaumatin II structural studies and, to date, data beyond atomic resolution had not been obtained. To identify the detailed structural properties explaining why thaumatin elicits a sweet taste, the structure of recombinant thaumatin II was determined at the resolution of 0.99 Å. Atomic resolution structural analysis with riding hydrogen atoms illustrated the differences in the direction of the side-chains more precisely and the electron density maps of the C-terminal regions were markedly improved. Though it had been suggested that the three consecutive glycine residues (G142-G143-G144) have highly flexible conformations, G143, the central glycine residue was successfully modelled in two conformations for the first time. Furthermore, the side chain r.m.s.d. values for two residues (R67 and R82) critical for sweetness exhibited substantially higher values, suggesting that these residues are highly disordered. These results demonstrated that the flexible conformations in two critical residues favoring their interaction with sweet taste receptors are prominent features of the intensely sweet taste of thaumatin.

  2. A microarray of ubiquitylated proteins for profiling deubiquitylase activity reveals the critical roles of both chain and substrate.

    Science.gov (United States)

    Loch, Christian M; Strickler, James E

    2012-11-01

    Substrate ubiquitylation is a reversible process critical to cellular homeostasis that is often dysregulated in many human pathologies including cancer and neurodegeneration. Elucidating the mechanistic details of this pathway could unlock a large store of information useful to the design of diagnostic and therapeutic interventions. Proteomic approaches to the questions at hand have generally utilized mass spectrometry (MS), which has been successful in identifying both ubiquitylation substrates and profiling pan-cellular chain linkages, but is generally unable to connect the two. Interacting partners of the deubiquitylating enzymes (DUBs) have also been reported by MS, although substrates of catalytically competent DUBs generally cannot be. Where they have been used towards the study of ubiquitylation, protein microarrays have usually functioned as platforms for the identification of substrates for specific E3 ubiquitin ligases. Here, we report on the first use of protein microarrays to identify substrates of DUBs, and in so doing demonstrate the first example of microarray proteomics involving multiple (i.e., distinct, sequential and opposing) enzymatic activities. This technique demonstrates the selectivity of DUBs for both substrate and type (mono- versus poly-) of ubiquitylation. This work shows that the vast majority of DUBs are monoubiquitylated in vitro, and are incapable of removing this modification from themselves. This work also underscores the critical role of utilizing both ubiquitin chains and substrates when attempting to characterize DUBs. This article is part of a Special Issue entitled: Ubiquitin Drug Discovery and Diagnostics.

  3. Crystal structures and molecular dynamics simulations of thermophilic malate dehydrogenase reveal critical loop motion for co-substrate binding.

    Science.gov (United States)

    Hung, Chih-Hung; Hwang, Tzann-Shun; Chang, Yu-Yung; Luo, Huei-Ru; Wu, Szu-Pei; Hsu, Chun-Hua

    2013-01-01

    Malate dehydrogenase (MDH) catalyzes the conversion of oxaloacetate and malate by using the NAD/NADH coenzyme system. The system is used as a conjugate for enzyme immunoassays of a wide variety of compounds, such as illegal drugs, drugs used in therapeutic applications and hormones. We elucidated the biochemical and structural features of MDH from Thermus thermophilus (TtMDH) for use in various biotechnological applications. The biochemical characterization of recombinant TtMDH revealed greatly increased activity above 60 °C and specific activity of about 2,600 U/mg with optimal temperature of 90 °C. Analysis of crystal structures of apo and NAD-bound forms of TtMDH revealed a slight movement of the binding loop and few structural elements around the co-substrate binding packet in the presence of NAD. The overall structures did not change much and retained all related positions, which agrees with the CD analyses. Further molecular dynamics (MD) simulation at higher temperatures were used to reconstruct structures from the crystal structure of TtMDH. Interestingly, at the simulated structure of 353 K, a large change occurred around the active site such that with increasing temperature, a mobile loop was closed to co-substrate binding region. From biochemical characterization, structural comparison and MD simulations, the thermal-induced conformational change of the co-substrate binding loop of TtMDH may contribute to the essential movement of the enzyme for admitting NAD and may benefit the enzyme's activity.

  4. Physiological and proteomic analyses of leaves from the halophyte Tangut Nitraria reveals diverse response pathways critical for high salinity tolerance

    Directory of Open Access Journals (Sweden)

    Tielong eCheng

    2015-02-01

    Full Text Available Soil salinization poses a serious threat to the environment and agricultural productivity worldwide. Studies on the physiological and molecular mechanisms of salinity tolerance in halophytic plants provide valuable information to enhance their salt tolerance. Tangut Nitraria is a widely distributed halophyte in saline–alkali soil in the northern areas of China. In this study, we used a proteomic approach to investigate the molecular pathways of the high salt tolerance of T. Nitraria. We analyzed the changes in biomass, photosynthesis, and redox-related enzyme activities in T. Nitraria leaves from plant seedlings treated with high salt concentration. Comparative proteomic analysis of the leaves revealed that the expression of 71 proteins was significantly altered after salinity treatments of T. Nitraria. These salinity-responsive proteins were mainly involved in photosynthesis, redox homeostasis, stress/defense, carbohydrate and energy metabolism, protein metabolism, signal transduction, and membrane transport. Results showed that the reduction of photosynthesis under salt stress was attributed to the down-regulation of the enzymes and proteins involved in the light reaction and Calvin cycle. Protein–protein interaction analysis revealed that the proteins involved in redox homeostasis, photosynthesis, and energy metabolism constructed two types of response networks to high salt stress. T. Nitraria plants developed diverse mechanisms for scavenging reactive oxygen species in their leaves to cope with stress induced by high salinity. This study provides important information regarding the salt tolerance of the halophyte T. Nitraria.

  5. Immune evasion activities of accessory proteins Vpu, Nef and Vif are conserved in acute and chronic HIV-1 infection.

    Science.gov (United States)

    Mlcochova, Petra; Apolonia, Luis; Kluge, Silvia F; Sridharan, Aishwarya; Kirchhoff, Frank; Malim, Michael H; Sauter, Daniel; Gupta, Ravindra K

    2015-08-01

    Heterosexual HIV-1 transmission has been identified as a genetic bottleneck and a single transmitted/founder (T/F) variant with reduced sensitivity to type I interferon initiates productive infection in most cases. We hypothesized that particularly active accessory protein(s) may confer T/F viruses with a selective advantage in establishing HIV infection. Thus, we tested vpu, vif and nef alleles from six T/F and six chronic (CC) viruses in assays for 9 immune evasion activities involving the counteraction of interferon-stimulated genes and modulation of ligands known to activate innate immune cells. All functions were highly conserved with no significant differences between T/F and CC viruses, suggesting that these accessory protein functions are important throughout the course of infection.

  6. Design of experiments reveals critical parameters for pilot-scale freeze-and-thaw processing of L-lactic dehydrogenase.

    Science.gov (United States)

    Roessl, Ulrich; Humi, Sebastian; Leitgeb, Stefan; Nidetzky, Bernd

    2015-09-01

    Freezing constitutes an important unit operation of biotechnological protein production. Effects of freeze-and-thaw (F/T) process parameters on stability and other quality attributes of the protein product are usually not well understood. Here a design of experiments (DoE) approach was used to characterize the F/T behavior of L-lactic dehydrogenase (LDH) in a 700-mL pilot-scale freeze container equipped with internal temperature and pH probes. In 24-hour experiments, target temperature between -10 and -38°C most strongly affected LDH stability whereby enzyme activity was retained best at the highest temperature of -10°C. Cooling profile and liquid fill volume also had significant effects on LDH stability and affected the protein aggregation significantly. Parameters of the thawing phase had a comparably small effect on LDH stability. Experiments in which the standard sodium phosphate buffer was exchanged by Tris-HCl and the non-ionic surfactant Tween 80 was added to the protein solution showed that pH shift during freezing and protein surface exposure were the main factors responsible for LDH instability at the lower freeze temperatures. Collectively, evidence is presented that supports the use of DoE-based systematic analysis at pilot scale in the identification of F/T process parameters critical for protein stability and in the development of suitable process control strategies.

  7. Molecular markers reveal spatially segregated cryptic species in a critically endangered fish, the common skate (Dipturus batis).

    Science.gov (United States)

    Griffiths, Andrew M; Sims, David W; Cotterell, Stephen P; El Nagar, Aliya; Ellis, Jim R; Lynghammar, Arve; McHugh, Matthew; Neat, Francis C; Pade, Nicolas G; Queiroz, Nuno; Serra-Pereira, Bárbara; Rapp, Toby; Wearmouth, Victoria J; Genner, Martin J

    2010-05-22

    Many sharks and skates are particularly vulnerable to overfishing because of their large size, slow growth, late maturity and low fecundity. In Europe dramatic population declines have taken place in common skate (Dipturus batis L.), one of the largest demersal fish in regional shelf seas, leading to extirpations from substantial parts of its former range. Here we report the discovery of cryptic species in common skate collected from the northeast Atlantic continental shelf. Data from nuclear microsatellite markers indicated two clearly distinct clades and phylogenetic analysis of mitochondrial DNA sequences demonstrated monophyly of each one of them. Capture locations showed evidence of strong spatial segregation, with one taxon occurring mainly in waters off the southern British Isles and around Rockall, while the other was restricted to more northerly shelf waters. These apparently cryptic species showed overlapping substrate and depth preferences, but distributional limits were closely related to temperature gradients, potentially indicating thermal limits to their distributions. This discovery of hidden diversity within a large, critically endangered marine vertebrate demonstrates how marine biodiversity can be underestimated, even in such a relatively well-studied and heavily exploited region.

  8. RNA Sequencing of Tumor-Associated Microglia Reveals Ccl5 as a Stromal Chemokine Critical for Neurofibromatosis-1 Glioma Growth

    Directory of Open Access Journals (Sweden)

    Anne C. Solga

    2015-10-01

    Full Text Available Solid cancers develop within a supportive microenvironment that promotes tumor formation and growth through the elaboration of mitogens and chemokines. Within these tumors, monocytes (macrophages and microglia represent rich sources of these stromal factors. Leveraging a genetically engineered mouse model of neurofibromatosis type 1 (NF1 low-grade brain tumor (optic glioma, we have previously demonstrated that microglia are essential for glioma formation and maintenance. To identify potential tumor-associated microglial factors that support glioma growth (gliomagens, we initiated a comprehensive large-scale discovery effort using optimized RNA-sequencing methods focused specifically on glioma-associated microglia. Candidate microglial gliomagens were prioritized to identify potential secreted or membrane-bound proteins, which were next validated by quantitative real-time polymerase chain reaction as well as by RNA fluorescence in situ hybridization following minocycline-mediated microglial inactivation in vivo. Using these selection criteria, chemokine (C-C motif ligand 5 (Ccl5 was identified as a chemokine highly expressed in genetically engineered Nf1 mouse optic gliomas relative to nonneoplastic optic nerves. As a candidate gliomagen, recombinant Ccl5 increased Nf1-deficient optic nerve astrocyte growth in vitro. Importantly, consistent with its critical role in maintaining tumor growth, treatment with Ccl5 neutralizing antibodies reduced Nf1 mouse optic glioma growth and improved retinal dysfunction in vivo. Collectively, these findings establish Ccl5 as an important microglial growth factor for low-grade glioma maintenance relevant to the development of future stroma-targeted brain tumor therapies.

  9. Analysis of core–periphery organization in protein contact networks reveals groups of structurally and functionally critical residues

    Indian Academy of Sciences (India)

    Arnold Emerson Isaac; Sitabhra Sinha

    2015-10-01

    The representation of proteins as networks of interacting amino acids, referred to as protein contact networks (PCN), and their subsequent analyses using graph theoretic tools, can provide novel insights into the key functional roles of specific groups of residues. We have characterized the networks corresponding to the native states of 66 proteins (belonging to different families) in terms of their core–periphery organization. The resulting hierarchical classification of the amino acid constituents of a protein arranges the residues into successive layers – having higher core order– with increasing connection density, ranging from a sparsely linked periphery to a densely intra-connected core (distinct from the earlier concept of protein core defined in terms of the three-dimensional geometry of the native state, which has least solvent accessibility). Our results show that residues in the inner cores are more conserved than those at the periphery. Underlining the functional importance of the network core, we see that the receptor sites for known ligand molecules of most proteins occur in the innermost core. Furthermore, the association of residues with structural pockets and cavities in binding or active sites increases with the core order. From mutation sensitivity analysis, we show that the probability of deleterious or intolerant mutations also increases with the core order. We also show that stabilization centre residues are in the innermost cores, suggesting that the network core is critically important in maintaining the structural stability of the protein. A publicly available Web resource for performing core–periphery analysis of any protein whose native state is known has been made available by us at http://www.imsc.res.in/ ~ sitabhra/proteinKcore/index.html.

  10. Molecular and morphological analysis of the critically endangered Fijian iguanas reveals cryptic diversity and a complex biogeographic history

    Science.gov (United States)

    Keogh, J.S.; Edwards, D.L.; Fisher, R.N.; Harlow, P.S.

    2008-01-01

    The Pacific iguanas of the Fijian and Tongan archipelagos are a biogeographic enigma in that their closest relatives are found only in the New World. They currently comprise two genera and four species of extinct and extant taxa. The two extant species, Brachylophus fasciatus from Fiji, Tonga, and Vanuatu and Brachylophus vitiensis from western Fiji, are of considerable conservation concern with B. vitiensis listed as critically endangered. A recent molecular study has shown that Brachylophus comprised three evolutionarily significant units. To test these conclusions and to reevaluate the phylogenetic and biogeographic relationships within Brachylophus, we generated an mtDNA dataset consisting of 1462 base pairs for 61 individuals from 13 islands, representing both Brachylophus species. Unweighted parsimony analyses and Bayesian analyses produced a well-resolved phylogenetic hypothesis supported by high bootstrap values and posterior probabilities within Brachylophus. Our data reject the monophyly of specimens previously believed to comprise B. fasciatus. Instead, our data demonstrate that living Brachylophus comprise three robust and well-supported clades that do not correspond to current taxonomy. One of these clades comprises B. fasciatus from the Lau group of Fiji and Tonga (type locality for B. fasciatus), while a second comprises putative B. fasciatus from the central regions of Fiji, which we refer to here as B. n. sp. Animals in this clade form the sister group to B. vitiensis rather than other B. fasciatus. We herein describe this clade as a new species of Brachylophus based on molecular and morphological data. With only one exception, every island is home to one or more unique haplotypes. We discuss alternative biogeographic hypotheses to explain their distribution in the Pacific and the difficulties of distinguishing these. Together, our molecular and taxonomic results have important implications for future conservation initiatives for the Pacific

  11. Structural Snapshots of an Engineered Cystathionine-γ-lyase Reveal the Critical Role of Electrostatic Interactions in the Active Site

    Energy Technology Data Exchange (ETDEWEB)

    Yan, Wupeng; Stone, Everett; Zhang, Yan Jessie

    2017-02-01

    Enzyme therapeutics that can degrade l-methionine (l-Met) are of great interest as numerous malignancies are exquisitely sensitive to l-Met depletion. To exhaust the pool of methionine in human serum, we previously engineered an l-Met-degrading enzyme based on the human cystathionine-γ-lyase scaffold (hCGL-NLV) to circumvent immunogenicity and stability issues observed in the preclinical application of bacterially derived methionine-γ-lyases. To gain further insights into the structure–activity relationships governing the chemistry of the hCGL-NLV lead molecule, we undertook a biophysical characterization campaign that captured crystal structures (2.2 Å) of hCGL-NLV with distinct reaction intermediates, including internal aldimine, substrate-bound, gem-diamine, and external aldimine forms. Curiously, an alternate form of hCGL-NLV that crystallized under higher-salt conditions revealed a locally unfolded active site, correlating with inhibition of activity as a function of ionic strength. Subsequent mutational and kinetic experiments pinpointed that a salt bridge between the phosphate of the essential cofactor pyridoxal 5'-phosphate (PLP) and residue R62 plays an important role in catalyzing β- and γ-eliminations. Our study suggests that solvent ions such as NaCl disrupt electrostatic interactions between R62 and PLP, decreasing catalytic efficiency.

  12. Crystal structures of ryanodine receptor SPRY1 and tandem-repeat domains reveal a critical FKBP12 binding determinant.

    Science.gov (United States)

    Yuchi, Zhiguang; Yuen, Siobhan M Wong King; Lau, Kelvin; Underhill, Ainsley Q; Cornea, Razvan L; Fessenden, James D; Van Petegem, Filip

    2015-08-06

    Ryanodine receptors (RyRs) form calcium release channels located in the membranes of the sarcoplasmic and endoplasmic reticulum. RyRs play a major role in excitation-contraction coupling and other Ca(2+)-dependent signalling events, and consist of several globular domains that together form a large assembly. Here we describe the crystal structures of the SPRY1 and tandem-repeat domains at 1.2-1.5 Å resolution, which reveal several structural elements not detected in recent cryo-EM reconstructions of RyRs. The cryo-EM studies disagree on the position of SPRY domains, which had been proposed based on homology modelling. Computational docking of the crystal structures, combined with FRET studies, show that the SPRY1 domain is located next to FK506-binding protein (FKBP). Molecular dynamics flexible fitting and mutagenesis experiments suggest a hydrophobic cluster within SPRY1 that is crucial for FKBP binding. A RyR1 disease mutation, N760D, appears to directly impact FKBP binding through interfering with SPRY1 folding.

  13. Lipidomic profiling of Saccharomyces cerevisiae and Zygosaccharomyces bailii reveals critical changes in lipid composition in response to acetic acid stress.

    Science.gov (United States)

    Lindberg, Lina; Santos, Aline Xs; Riezman, Howard; Olsson, Lisbeth; Bettiga, Maurizio

    2013-01-01

    When using microorganisms as cell factories in the production of bio-based fuels or chemicals from lignocellulosic hydrolysate, inhibitory concentrations of acetic acid, released from the biomass, reduce the production rate. The undissociated form of acetic acid enters the cell by passive diffusion across the lipid bilayer, mediating toxic effects inside the cell. In order to elucidate a possible link between lipid composition and acetic acid stress, the present study presents detailed lipidomic profiling of the major lipid species found in the plasma membrane, including glycerophospholipids, sphingolipids and sterols, in Saccharomyces cerevisiae (CEN.PK 113_7D) and Zygosaccharomyces bailii (CBS7555) cultured with acetic acid. Detailed physiological characterization of the response of the two yeasts to acetic acid has also been performed in aerobic batch cultivations using bioreactors. Physiological characterization revealed, as expected, that Z. bailii is more tolerant to acetic acid than S. cerevisiae. Z. bailii grew at acetic acid concentrations above 24 g L(-1), while limited growth of S. cerevisiae was observed after 11 h when cultured with only 12 g L(-1) acetic acid. Detailed lipidomic profiling using electrospray ionization, multiple-reaction-monitoring mass spectrometry (ESI-MRM-MS) showed remarkable changes in the glycerophospholipid composition of Z. bailii, including an increase in saturated glycerophospholipids and considerable increases in complex sphingolipids in both S. cerevisiae (IPC 6.2×, MIPC 9.1×, M(IP)2C 2.2×) and Z. bailii (IPC 4.9×, MIPC 2.7×, M(IP)2C 2.7×), when cultured with acetic acid. In addition, the basal level of complex sphingolipids was significantly higher in Z. bailii than in S. cerevisiae, further emphasizing the proposed link between lipid saturation, high sphingolipid levels and acetic acid tolerance. The results also suggest that acetic acid tolerance is associated with the ability of a given strain to generate large

  14. Lipidomic Profiling of Saccharomyces cerevisiae and Zygosaccharomyces bailii Reveals Critical Changes in Lipid Composition in Response to Acetic Acid Stress

    Science.gov (United States)

    Riezman, Howard; Olsson, Lisbeth; Bettiga, Maurizio

    2013-01-01

    When using microorganisms as cell factories in the production of bio-based fuels or chemicals from lignocellulosic hydrolysate, inhibitory concentrations of acetic acid, released from the biomass, reduce the production rate. The undissociated form of acetic acid enters the cell by passive diffusion across the lipid bilayer, mediating toxic effects inside the cell. In order to elucidate a possible link between lipid composition and acetic acid stress, the present study presents detailed lipidomic profiling of the major lipid species found in the plasma membrane, including glycerophospholipids, sphingolipids and sterols, in Saccharomyces cerevisiae (CEN.PK 113_7D) and Zygosaccharomyces bailii (CBS7555) cultured with acetic acid. Detailed physiological characterization of the response of the two yeasts to acetic acid has also been performed in aerobic batch cultivations using bioreactors. Physiological characterization revealed, as expected, that Z. bailii is more tolerant to acetic acid than S. cerevisiae. Z. bailii grew at acetic acid concentrations above 24 g L−1, while limited growth of S. cerevisiae was observed after 11 h when cultured with only 12 g L−1 acetic acid. Detailed lipidomic profiling using electrospray ionization, multiple-reaction-monitoring mass spectrometry (ESI-MRM-MS) showed remarkable changes in the glycerophospholipid composition of Z. bailii, including an increase in saturated glycerophospholipids and considerable increases in complex sphingolipids in both S. cerevisiae (IPC 6.2×, MIPC 9.1×, M(IP)2C 2.2×) and Z. bailii (IPC 4.9×, MIPC 2.7×, M(IP)2C 2.7×), when cultured with acetic acid. In addition, the basal level of complex sphingolipids was significantly higher in Z. bailii than in S. cerevisiae, further emphasizing the proposed link between lipid saturation, high sphingolipid levels and acetic acid tolerance. The results also suggest that acetic acid tolerance is associated with the ability of a given strain to generate large

  15. Lipidomic profiling of Saccharomyces cerevisiae and Zygosaccharomyces bailii reveals critical changes in lipid composition in response to acetic acid stress.

    Directory of Open Access Journals (Sweden)

    Lina Lindberg

    Full Text Available When using microorganisms as cell factories in the production of bio-based fuels or chemicals from lignocellulosic hydrolysate, inhibitory concentrations of acetic acid, released from the biomass, reduce the production rate. The undissociated form of acetic acid enters the cell by passive diffusion across the lipid bilayer, mediating toxic effects inside the cell. In order to elucidate a possible link between lipid composition and acetic acid stress, the present study presents detailed lipidomic profiling of the major lipid species found in the plasma membrane, including glycerophospholipids, sphingolipids and sterols, in Saccharomyces cerevisiae (CEN.PK 113_7D and Zygosaccharomyces bailii (CBS7555 cultured with acetic acid. Detailed physiological characterization of the response of the two yeasts to acetic acid has also been performed in aerobic batch cultivations using bioreactors. Physiological characterization revealed, as expected, that Z. bailii is more tolerant to acetic acid than S. cerevisiae. Z. bailii grew at acetic acid concentrations above 24 g L(-1, while limited growth of S. cerevisiae was observed after 11 h when cultured with only 12 g L(-1 acetic acid. Detailed lipidomic profiling using electrospray ionization, multiple-reaction-monitoring mass spectrometry (ESI-MRM-MS showed remarkable changes in the glycerophospholipid composition of Z. bailii, including an increase in saturated glycerophospholipids and considerable increases in complex sphingolipids in both S. cerevisiae (IPC 6.2×, MIPC 9.1×, M(IP2C 2.2× and Z. bailii (IPC 4.9×, MIPC 2.7×, M(IP2C 2.7×, when cultured with acetic acid. In addition, the basal level of complex sphingolipids was significantly higher in Z. bailii than in S. cerevisiae, further emphasizing the proposed link between lipid saturation, high sphingolipid levels and acetic acid tolerance. The results also suggest that acetic acid tolerance is associated with the ability of a given strain to

  16. TE-domestication and horizontal transfer in a putative Nef-AP1mu mimic of HLA-A cytoplasmic domain re-trafficking.

    Science.gov (United States)

    Murray, Joseph S; Murray, Elaina H

    2016-01-01

    Genes of the major histocompatibility complex (MHC; also called HLA in human) are polymorphic elements in the genomes of sharks to humans. Class-I and class-II MHC loci appear responsible for much of the genetic linkage to myriad disease states via the capacity to bind short (~8-15 a.a.) peptides of a given pathogen's proteome, or in some cases, the altered proteomes of cancerous cells, and even (in autoimmunity) certain nominal 'self' peptides (Janeway, 2004).(1) Unfortunately, little is known about how the canonical structure of the MHC-I/-II peptide-presenting gene evolved, particularly since beyond ~500 Mya (sharks) no paralogs exist.(2,3) We previously reported that HLA-A isotype alleles with the α1-helix, R65 motif, are wide-spread in phylogeny, but that the α 2-helix, H151R motif, has apparently segregated out of most species. Surprisingly, an uncharacterized orf in T. syrichta (Loc-103275158) encoded R151, but within a truncated A-23 like gene containing 5'- and 3'- footprints of the transposon (TE), tigger-1; the extant tarsier A-23 allele is totally missing exon-3 and part-of exon-4; together, suggesting TE-mediated inactivation of an intact/ancestral A-23 allele (Murray, 2015a).(4) The unique Loc-103275158 orf encodes a putative 15-exon transcript with no apparent paralogs throughout phylogeny. However, an HLA-A11 like gene in M. leucophaeus with a shortened C-terminal domain, and an HLA-A like orf in C. atys with two linked α1/α2/α3 domains, both contain a second transmembrane segment, which is conserved in Loc-103275158. Thus, we could model the putative protein with its Nef-like tail domain docked to its MHC-I like α3 domain (i.e., on the same side of a membrane). This modeled tertiary structure is strikingly similar to the solved structure of the Nef:MHC-I CD:AP1mu transporter (Jia, 2012).(5) Nef:AP1mu binds the CD of MHC-I in trafficking MHC-I away from the trans-golgi and into the endocytic pathway in HIV-1 infected cells. The CD loop of the

  17. Differential Regulation of NF-κB-Mediated Proviral and Antiviral Host Gene Expression by Primate Lentiviral Nef and Vpu Proteins

    Directory of Open Access Journals (Sweden)

    Daniel Sauter

    2015-02-01

    Full Text Available NF-κB is essential for effective transcription of primate lentiviral genomes and also activates antiviral host genes. Here, we show that the early protein Nef of most primate lentiviruses enhances NF-κB activation. In contrast, the late protein Vpu of HIV-1 and its simian precursors inhibits activation of NF-κB, even in the presence of Nef. Although this effect of Vpu did not correlate with its ability to interact with β-TrCP, it involved the stabilization of IκB and reduced nuclear translocation of p65. Interestingly, however, Vpu did not affect casein kinase II-mediated phosphorylation of p65. Lack of Vpu was associated with increased NF-κB activation and induction of interferon and interferon-stimulated genes (ISGs in HIV-1-infected T cells. Thus, HIV-1 and its simian precursors employ Nef to boost NF-κB activation early during the viral life cycle to initiate proviral transcription, while Vpu is used to downmodulate NF-κB-dependent expression of ISGs at later stages.

  18. Identification of new and unusual rev and nef transcripts expressed by an HIV type 1 primary isolate.

    Science.gov (United States)

    Vega, Yolanda; Delgado, Elena; Carrera, Cristina; Nebreda, Paloma; Fernández-García, Aurora; Cuevas, María Teresa; Pérez-Álvarez, Lucía; Thomson, Michael M

    2013-07-01

    We analyzed RNA splice site usage in three HIV-1 subtype B primary isolates through reverse transcriptase polymerase chain reaction (RT-PCR) amplification of spliced RNAs using a fluorescently labeled primer, with computerized size determination and quantification of PCR products, which were also identified by clone sequencing. In one isolate, P2149-3, unusual and unreported spliced transcripts were detected. This isolate preferentially used for rev RNA generation a 3' splice site (3'ss) located five nucleotides upstream of A4a, previously identified only in a T cell line-adapted virus and in a group O isolate, and designated A4d. P2149-3 also used an unreported 3'ss for rev RNA generation, designated A4h, located 20 nucleotides upstream of 3'ss A4c. Additionally, unusual nef RNAs using 3'ss A5a and A7a and with exon composition 1.3.7 were identified. The identification of several unusual and unreported spliced transcripts in an HIV-1 primary isolate suggests a greater diversity of splice site usage in HIV-1 than previously appreciated.

  19. Plant-based strategies aimed at expressing HIV antigens and neutralizing antibodies at high levels. Nef as a case study.

    Science.gov (United States)

    Marusic, Carla; Vitale, Alessandro; Pedrazzini, Emanuela; Donini, Marcello; Frigerio, Lorenzo; Bock, Ralph; Dix, Philip J; McCabe, Matthew S; Bellucci, Michele; Benvenuto, Eugenio

    2009-08-01

    The first evidence that plants represent a valid, safe and cost-effective alternative to traditional expression systems for large-scale production of antigens and antibodies was described more than 10 years ago. Since then, considerable improvements have been made to increase the yield of plant-produced proteins. These include the use of signal sequences to target proteins to different cellular compartments, plastid transformation to achieve high transgene dosage, codon usage optimization to boost gene expression, and protein fusions to improve recombinant protein stability and accumulation. Thus, several HIV/SIV antigens and neutralizing anti-HIV antibodies have recently been successfully expressed in plants by stable nuclear or plastid transformation, and by transient expression systems based on plant virus vectors or Agrobacterium-mediated infection. The current article gives an overview of plant expressed HIV antigens and antibodies and provides an account of the use of different strategies aimed at increasing the expression of the accessory multifunctional HIV-1 Nef protein in transgenic plants.

  20. Genetic Diversity and Population Structure of the Critically Endangered Yangtze Finless Porpoise (Neophocaena asiaeorientalis asiaeorientalis as Revealed by Mitochondrial and Microsatellite DNA

    Directory of Open Access Journals (Sweden)

    Minmin Chen

    2014-06-01

    Full Text Available Ecological surveys have indicated that the population of the critically endangered Yangtze finless porpoise (YFP, Neophocaena asiaeorientalis asiaeorientalis is becoming increasingly small and fragmented, and will be at high risk of extinction in the near future. Genetic conservation of this population will be an important component of the long-term conservation effort. We used a 597 base pair mitochondrial DNA (mtDNA control region and 11 microsatellite loci to analyze the genetic diversity and population structure of the YFP. The analysis of both mtDNA and microsatellite loci suggested that the genetic diversity of the YFP will possibly decrease in the future if the population keeps declining at a rapid rate, even though these two types of markers revealed different levels of genetic diversity. In addition, mtDNA revealed strong genetic differentiation between one local population, Xingchang–Shishou (XCSS, and the other five downstream local populations; furthermore, microsatellite DNA unveiled fine but significant genetic differentiation between three of the local populations (not only XCSS but also Poyang Lake (PY and Tongling (TL and the other local populations. With an increasing number of distribution gaps appearing in the Yangtze main steam, the genetic differentiation of local populations will likely intensify in the future. The YFP is becoming a genetically fragmented population. Therefore, we recommend attention should be paid to the genetic conservation of the YFP.

  1. Critical Role of CD2 Co-stimulation in Adaptive Natural Killer Cell Responses Revealed in NKG2C-Deficient Humans

    Directory of Open Access Journals (Sweden)

    Lisa L. Liu

    2016-05-01

    Full Text Available Infection by human cytomegalovirus (HCMV leads to NKG2C-driven expansion of adaptive natural killer (NK cells, contributing to host defense. However, approximately 4% of all humans carry a homozygous deletion of the gene that encodes NKG2C (NKG2C−/−. Assessment of NK cell repertoires in 60 NKG2C−/− donors revealed a broad range of NK cell populations displaying characteristic footprints of adaptive NK cells, including a terminally differentiated phenotype, functional reprogramming, and epigenetic remodeling of the interferon (IFN-γ promoter. We found that both NKG2C− and NKG2C+ adaptive NK cells expressed high levels of CD2, which synergistically enhanced ERK and S6RP phosphorylation following CD16 ligation. Notably, CD2 co-stimulation was critical for the ability of adaptive NK cells to respond to antibody-coated target cells. These results reveal an unexpected redundancy in the human NK cell response to HCMV and suggest that CD2 provides “signal 2” in antibody-driven adaptive NK cell responses.

  2. Event-related repetitive TMS reveals distinct, critical roles for right OFA and bilateral posterior STS in judging the sex and trustworthiness of faces.

    Science.gov (United States)

    Dzhelyova, Milena P; Ellison, Amanda; Atkinson, Anthony P

    2011-10-01

    Judging the sex of faces relies on cues related to facial morphology and spatial relations between features, whereas judging the trustworthiness of faces relies on both structural and expressive cues that signal affective valence. The right occipital face area (OFA) processes structural cues and has been associated with sex judgments, whereas the posterior STS processes changeable facial cues related to muscle movements and is activated when observers judge trustworthiness. It is commonly supposed that the STS receives inputs from the OFA, yet it is unknown whether these regions have functionally dissociable, critical roles in sex and trustworthiness judgments. We addressed this issue using event-related, fMRI-guided repetitive transcranial magnetic stimulation (rTMS). Twelve healthy volunteers judged the sex of individually presented faces and, in a separate session, whether those same faces were trustworthy or not. Relative to sham stimulation, RTs were significantly longer for sex judgments when rTMS was delivered over the right OFA but not the right or left STS, and for trustworthiness judgments on male but not female faces when rTMS was delivered over the right STS or left STS but not the right OFA. Nonetheless, an analysis of the RT distributions revealed a possible critical role also for the right OFA in trustworthiness judgments, limited to faces with longer RTs, perhaps reflecting the later, ancillary use of structural cues related to the sex of the face. On the whole, our findings provide evidence that evaluations of the trustworthiness and sex of faces rely on functionally dissociable cortical regions.

  3. Increased breadth and depth of cytotoxic T lymphocytes responses against HIV-1-B Nef by inclusion of epitope variant sequences.

    Directory of Open Access Journals (Sweden)

    Morgane Rolland

    Full Text Available Different vaccine approaches cope with HIV-1 diversity, ranging from centralized(1-4 to variability-encompassing(5-7 antigens. For all these strategies, a concern remains: how does HIV-1 diversity impact epitope recognition by the immune system? We studied the relationship between HIV-1 diversity and CD8(+ T Lymphocytes (CTL targeting of HIV-1 subtype B Nef using 944 peptides (10-mers overlapping by nine amino acids (AA that corresponded to consensus peptides and their most common variants in the HIV-1-B virus population. IFN-γ ELISpot assays were performed using freshly isolated PBMC from 26 HIV-1-infected persons. Three hundred and fifty peptides elicited a response in at least one individual. Individuals targeted a median of 7 discrete regions. Overall, 33% of responses were directed against viral variants but not elicited against consensus-based test peptides. However, there was no significant relationship between the frequency of a 10-mer in the viral population and either its frequency of recognition (Spearman's correlation coefficient ρ = 0.24 or the magnitude of the responses (ρ = 0.16. We found that peptides with a single mutation compared to the consensus were likely to be recognized (especially if the change was conservative and to elicit responses of similar magnitude as the consensus peptide. Our results indicate that cross-reactivity between rare and frequent variants is likely to play a role in the expansion of CTL responses, and that maximizing antigenic diversity in a vaccine may increase the breadth and depth of CTL responses. However, since there are few obvious preferred pathways to virologic escape, the diversity that may be required to block all potential escape pathways may be too large for a realistic vaccine to accommodate. Furthermore, since peptides were not recognized based on their frequency in the population, it remains unclear by which mechanisms variability-inclusive antigens (i.e., constructs enriched with

  4. Persistence of attenuated HIV-1 rev alleles in an epidemiologically linked cohort of long-term survivors infected with nef-deleted virus

    Directory of Open Access Journals (Sweden)

    Wesselingh Steven L

    2007-07-01

    Full Text Available Abstract Background The Sydney blood bank cohort (SBBC of long-term survivors consists of multiple individuals infected with nef-deleted, attenuated strains of human immunodeficiency virus type 1 (HIV-1. Although the cohort members have experienced differing clinical courses and now comprise slow progressors (SP as well as long-term nonprogressors (LTNP, longitudinal analysis of nef/long-terminal repeat (LTR sequences demonstrated convergent nef/LTR sequence evolution in SBBC SP and LTNP. Thus, the in vivo pathogenicity of attenuated HIV-1 strains harboured by SBBC members is dictated by factors other than nef/LTR. Therefore, to determine whether defects in other viral genes contribute to attenuation of these HIV-1 strains, we characterized dominant HIV-1 rev alleles that persisted in 4 SBBC subjects; C18, C64, C98 and D36. Results The ability of Rev derived from D36 and C64 to bind the Rev responsive element (RRE in RNA binding assays was reduced by approximately 90% compared to Rev derived from HIV-1NL4-3, C18 or C98. D36 Rev also had a 50–60% reduction in ability to express Rev-dependent reporter constructs in mammalian cells. In contrast, C64 Rev had only marginally decreased Rev function despite attenuated RRE binding. In D36 and C64, attenuated RRE binding was associated with rare amino acid changes at 3 highly conserved residues; Gln to Pro at position 74 immediately N-terminal to the Rev activation domain, and Val to Leu and Ser to Pro at positions 104 and 106 at the Rev C-terminus, respectively. In D36, reduced Rev function was mapped to an unusual 13 amino acid extension at the Rev C-terminus. Conclusion These findings provide new genetic and mechanistic insights important for Rev function, and suggest that Rev function, not Rev/RRE binding may be rate limiting for HIV-1 replication. In addition, attenuated rev alleles may contribute to viral attenuation and long-term survival of HIV-1 infection in a subset of SBBC members.

  5. Crystal Structure of Acivicin-Inhibited [gamma]-Glutamyltranspeptidase Reveals Critical Roles for Its C-Terminus in Autoprocessing and Catalysis

    Energy Technology Data Exchange (ETDEWEB)

    Williams, Kristin; Cullati, Sierra; Sand, Aaron; Biterova, Ekaterina I.; Barycki, Joseph J.; (UNL)

    2009-03-27

    Helicobacter pylori {gamma}-glutamyltranspeptidase (HpGT) is a general {gamma}-glutamyl hydrolase and a demonstrated virulence factor. The enzyme confers a growth advantage to the bacterium, providing essential amino acid precursors by initiating the degradation of extracellular glutathione and glutamine. HpGT is a member of the N-terminal nucleophile (Ntn) hydrolase superfamily and undergoes autoprocessing to generate the active form of the enzyme. Acivicin is a widely used {gamma}-glutamyltranspeptidase inhibitor that covalently modifies the enzyme, but its precise mechanism of action remains unclear. The time-dependent inactivation of HpGT exhibits a hyperbolic dependence on acivicin concentration with k{sub max} = 0.033 {+-} 0.006 s{sup -1} and K{sub I} = 19.7 {+-} 7.2 {micro}M. Structure determination of acivicin-modified HpGT (1.7 {angstrom}; R{sub factor} = 17.9%; R{sub free} = 20.8%) demonstrates that acivicin is accommodated within the {gamma}-glutamyl binding pocket of the enzyme. The hydroxyl group of Thr 380, the catalytic nucleophile in the autoprocessing and enzymatic reactions, displaces chloride from the acivicin ring to form the covalently linked complex. Within the acivicin-modified HpGT structure, the C-terminus of the protein becomes ordered with Phe 567 positioned over the active site. Substitution or deletion of Phe 567 leads to a >10-fold reduction in enzymatic activity, underscoring its importance in catalysis. The mobile C-terminus is positioned by several electrostatic interactions within the C-terminal region, most notably a salt bridge between Arg 475 and Glu 566. Mutational analysis reveals that Arg 475 is critical for the proper placement of the C-terminal region, the Tyr 433 containing loop, and the proposed oxyanion hole.

  6. NEF-HIV-1 incrementa la expresión del factor de transcripción FOXP3 en monocitos humanos

    Directory of Open Access Journals (Sweden)

    Juan Camilo Valencia

    2013-12-01

    Full Text Available Un mecanismo utilizado por el VIH para evadir la respuesta inmune es favoreciendo una respuesta reguladora al incrementar las Células T Reguladoras (TREG durante la infección, estas células expresan Foxp3, un factor de transcripción asociado con actividad supresora, cuya presencia ha sido confirmada en monocitos y macrófagos. Siendo los monocitos/macrófagos susceptibles a la infección VIH-1, y uno de los principales reservorios virales, se evaluó el papel de Nef (una de las principales proteínas reguladoras del VIH, en la expresión de Foxp3 en monocitos humanos. Nef-VIH-1 fue expresada en E. coli y la proteína purificada se expuso a monocitos de sangre periférica de individuos seronegativos. La expresión de Foxp3 fue analizada mediante citometría de flujo y western-blot.

  7. IL-12 and GM-CSF in DNA/MVA immunizations against HIV-1 CRF12_BF Nef induced T-cell responses with an enhanced magnitude, breadth and quality.

    Directory of Open Access Journals (Sweden)

    Ana María Rodríguez

    Full Text Available In Argentina, the HIV epidemic is characterized by the co-circulation of subtype B and BF recombinant viral variants. Nef is an HIV protein highly variable among subtypes, making it a good tool to study the impact of HIV variability in the vaccine design setting. We have previously reported a specific cellular response against NefBF with low cross-reactivity to NefB in mice. The aim of this work was to analyze whether the co-administration of IL-12 and GM-CSF, using DNA and MVA vaccine vectors, could improve the final cellular response induced. Mice received three DNA priming doses of a plasmid that express NefBF plus DNAs expressing IL-12 and/or GM-CSF. Afterwards, all the groups were boosted with a MVAnefBF dose. The highest increase in the magnitude of the NefBF response, compared to that induced in the control was found in the IL-12 group. Importantly, a response with higher breadth was detected in groups which received IL-12 or GM-CSF, evidenced as an increased frequency of recognition of homologous (BF and heterologous (B Nef peptides, as well as a higher number of other Nef peptide pools representing different viral subtypes. However, these improvements were lost when both DNA cytokines were simultaneously administered, as the response was focused against the immunodominant peptide with a detrimental response towards subdominant epitopes. The pattern of cytokines secreted and the specific-T-cell proliferative capacity were improved in IL-12 and IL-12+GM-CSF groups. Importantly IL-12 generated a significant higher T-cell avidity against a B heterologous peptide.This study indicates that the incorporation of DNA expressing IL-12 in DNA/MVA schemes produced the best results in terms of improvements of T-cell-response key properties such as breadth, cross-reactivity and quality (avidity and pattern of cytokines secreted. These relevant results contribute to the design of strategies aimed to induce T-cell responses against HIV antigens with

  8. Prenatal exposure of mice to the human liver carcinogen Aflatoxin B1 reveals a critical window of susceptibility to genetic change

    OpenAIRE

    2014-01-01

    It has become axiomatic that critical windows of susceptibility to genotoxins exist and that genetic damage in utero may be a trigger for later life cancers. Data supporting this critical window hypothesis are remarkably few. This study provides a quantitative bridge between DNA damage by the liver carcinogen aflatoxin B1 (AFB1) during prenatal development and the risk of later life genetic disease. AFB1 was given to pregnant C57BL/6J mice, carrying F1 gestation day 14 (GD14) embryos of the B...

  9. A Biochemical/Biophysical Assay Dyad for HTS-Compatible Triaging of Inhibitors of the HIV-1 Nef/Hck SH3 Interaction

    KAUST Repository

    Breuer, Sebastian

    2013-07-26

    The current treatment regimens for HIV include over 20 anti-retrovirals. However, adverse drug effects and the emergence of drug resistance necessitates the continued improvement of the existing drug classes as well as the development of novel drugs that target as yet therapeutically unexploited viral and cellular pathways. Here we demonstrate a strategy for the discovery of protein-protein interaction inhibitors of the viral pathogenicity factor HIV-1 Nef and its interaction with the host factor SH3. A combination of a time-resolved fluorescence resonance energy resonance energy transfer-based assay and a label-free resonant waveguide grating-based assay was optimized for high-throughput screening formats.

  10. Tetherin does not significantly restrict dendritic cell-mediated HIV-1 transmission and its expression is upregulated by newly synthesized HIV-1 Nef

    Directory of Open Access Journals (Sweden)

    Wu Li

    2011-04-01

    Full Text Available Abstract Background Dendritic cells (DCs are among the first cells to encounter HIV-1 and play important roles in viral transmission and pathogenesis. Immature DCs allow productive HIV-1 replication and long-term viral dissemination. The pro-inflammatory factor lipopolysaccharide (LPS induces DC maturation and enhances the efficiency of DC-mediated HIV-1 transmission. Type I interferon (IFN partially inhibits HIV-1 replication and cell-cell transmission in CD4+ T cells and macrophages. Tetherin is a type I IFN-inducible restriction factor that blocks HIV-1 release and modulates CD4+ T cell-mediated cell-to-cell transmission of HIV-1. However, the role of type I IFN and tetherin in HIV-1 infection of DCs and DC-mediated viral transmission remains unknown. Results We demonstrated that IFN-alpha (IFNα-induced mature DCs restricted HIV-1 replication and trans-infection of CD4+ T cells. Tetherin expression in monocyte-derived immature DCs was undetectable or very low. High levels of tetherin were transiently expressed in LPS- and IFNα-induced mature DCs, while HIV-1 localized into distinct patches in these DCs. Knockdown of induced tetherin in LPS- or IFNα-matured DCs modestly enhanced HIV-1 transmission to CD4+ T cells, but had no significant effect on wild-type HIV-1 replication in mature DCs. Intriguingly, we found that HIV-1 replication in immature DCs induced significant tetherin expression in a Nef-dependent manner. Conclusions The restriction of HIV-1 replication and transmission in IFNα-induced mature DCs indicates a potent anti-HIV-1 response; however, high levels of tetherin induced in mature DCs cannot significantly restrict wild-type HIV-1 release and DC-mediated HIV-1 transmission. Nef-dependent tetherin induction in HIV-1-infected immature DCs suggests an innate immune response of DCs to HIV-1 infection.

  11. Conditional IL-4/IL-13-deficient mice reveal a critical role of innate immune cells for protective immunity against gastrointestinal helminths.

    Science.gov (United States)

    Oeser, K; Schwartz, C; Voehringer, D

    2015-05-01

    Approximately one-third of the world population is infected with gastrointestinal helminths. Studies in mouse models have demonstrated that the cytokines interleukin (IL)-4 and IL-13 are essential for worm expulsion, but the critical cellular source of these cytokines is poorly defined. Here, we compared the immune response to Nippostrongylus brasiliensis in wild-type, T cell-specific IL-4/IL-13-deficient and general IL-4/IL-13-deficient mice. We show that T cell-derived IL-4/IL-13 promoted T helper 2 (Th2) polarization in a paracrine manner, differentiation of alternatively activated macrophages, and tissue recruitment of innate effector cells. However, innate IL-4/IL-13 played the critical role for induction of goblet cell hyperplasia and secretion of effector molecules like Mucin5ac and RELMβ in the small intestine. Surprisingly, T cell-specific IL-4/IL-13-deficient and wild-type mice cleared the parasite with comparable efficiency, whereas IL-4/IL-13-deficient mice showed impaired expulsion. These findings demonstrate that IL-4/IL-13 produced by cells of the innate immune system is required and sufficient to initiate effective type 2 immune responses resulting in protective immunity against N. brasiliensis.

  12. El dominio de anclaje a la membrana de NEF-HIV-1 es crítico en la expresión de FOXP3 en Monocitos humanos

    Directory of Open Access Journals (Sweden)

    Juan Camilo Valencia

    2013-12-01

    Full Text Available Introducción: El VIH, durante la infección, favorece una respuesta reguladora al incrementar las Células T Reguladoras (TREG para evadir la respuesta inmune, estas células expresan Foxp3, un factor de transcripción asociado con actividad supresora, cuya presencia ha sido confirmada en monocitos y macrófagos; Siendo estos susceptibles a la infección VIH-1, y uno de los principales reservorios virales, se evaluó el papel de los diferentes dominios de Nef, en la expresión de Foxp3 en monocitos humanos. Diferentes formas truncadas de Nef-VIH-1 fueron expresadas en E. coli (el dominio de anclaje a la membrana, SH3, PAK1, de dimerización y PAK2 y expuestas a monocitos humanos.

  13. Caracterização da interação de proteínas celulares envolvidas na degradação de Cd4 mediada por diferentes alelos da proteína Nef do Hiv-1

    OpenAIRE

    Gondim, Marcos Vinícius Pereira

    2011-01-01

    A diminuição da expressão do receptor CD4 na superfície das células infectadas é um dos mais importantes eventos durante a infecção pelo HIV-1. Foi observada uma clara relação entre a degradação do receptor viral, o aumento da infectuosidade e a replicação viral, sugerindo a participação deste fenômeno na patogênese e progressão da infecção. Para vencer os efeitos negativos exercidos pela expressão da molécula CD4, o HIV possui três proteínas: Nef, Vpu e Env, sendo Nef a proteína mais relevan...

  14. Toward Revealing the Critical Role of Perovskite Coverage in Highly Efficient Electron-Transport Layer-Free Perovskite Solar Cells: An Energy Band and Equivalent Circuit Model Perspective.

    Science.gov (United States)

    Huang, Like; Xu, Jie; Sun, Xiaoxiang; Du, Yangyang; Cai, Hongkun; Ni, Jian; Li, Juan; Hu, Ziyang; Zhang, Jianjun

    2016-04-20

    Currently, most efficient perovskite solar cells (PVKSCs) with a p-i-n structure require simultaneously electron transport layers (ETLs) and hole transport layers (HTLs) to help collecting photogenerated electrons and holes for obtaining high performance. ETL free planar PVKSC is a relatively new and simple structured solar cell that gets rid of the complex and high temperature required ETL (such as compact and mesoporous TiO2). Here, we demonstrate the critical role of high coverage of perovskite in efficient ETL free PVKSCs from an energy band and equivalent circuit model perspective. From an electrical point of view, we confirmed that the low coverage of perovskite does cause localized short circuit of the device. With coverage optimization, a planar p-i-n(++) device with a power conversion efficiency of over 11% was achieved, implying that the ETL layer may not be necessary for an efficient device as long as the perovskite coverage is approaching 100%.

  15. La charpente de la nef de la cathédrale de Bourges The structure of the nave in Bourges cathedral

    Directory of Open Access Journals (Sweden)

    Frédéric Epaud

    2012-03-01

    Full Text Available La cathédrale gothique de Bourges a été édifiée en deux campagnes avec le chœur, de 1195 à 1214, puis la nef, de 1225 à 1255, après 10 ans d’interruption du chantier. Ses charpentes d’origine ont été en partie reconstruites sur l’ensemble des collatéraux, suite à un incendie en 1559, et sur la partie médiane du grand vaisseau, entre 1747 et 1754, du fait de la suppression du faux transept et de sa flèche. Le relevé archéologique de la charpente gothique subsistante sur la nef, et l’analyse dendrochronologique de ses bois démontrent que les abattages se sont étalés de 1230 à 1257, avec deux grandes campagnes de coupes de bois entre 1240 et 1244 et en automne-hiver 1254-1255, juste avant la mise en œuvre et le levage de la charpente en 1256 ou peu après. Ces coupes successives supposent un approvisionnement du chantier par des donations de bois ou de parcelles forestières exploitées aussitôt, obligeant à un stockage des bois dans l’attente du chantier. La charpente à chevrons-formant-fermes a été exécutée selon un premier projet qui a été modifié en cours de réalisation pour y insérer un dispositif de contreventement longitudinal, avec une nouvelle structure des fermes principales permettant de le recevoir. Ce contreventement axial est doublé par un second situé dans le plan des chevrons, attesté pour la première fois par la dendrochronologie du milieu du xiiie s. En 1262, les travées orientales de la charpente de la nef sont reprises pour permettre le rajout d’une flèche en bois et d’un faux-transept à l’aplomb de la quatrième travée de voûtes.The Gothic cathedral of Bourges was built in two phases, with the chancel from 1195 to 1214 then the nave from 1225 to 1255, after a 10-year interruption to the building work. The original roof structure was in part reconstructed on all the sides, following a fire in 1559, and on the middle part of the great nave, between 1747 and 1754, due to

  16. Short Communication: HIV-1 Variants That Use Mouse CCR5 Reveal Critical Interactions of gp120's V3 Crown with CCR5 Extracellular Loop 1.

    Science.gov (United States)

    Platt, Emily J; Durnin, James P; Kabat, David

    2015-10-01

    The CCR5 coreceptor amino terminus and extracellular (ECL) loops 1 and 2 have been implicated in HIV-1 infections, with species differences in these regions inhibiting zoonoses. Interactions of gp120 with CD4 and CCR5 reduce constraints on metastable envelope subunit gp41, enabling gp41 conformational changes needed for infection. We previously selected HIV-1JRCSF variants that efficiently use CCR5(Δ18) with a deleted amino terminus or CCR5(HHMH) with ECL2 from an NIH/Swiss mouse. Unexpectedly, the adaptive gp120 mutations were nearly identical, suggesting that they function by weakening gp120's grip on gp41 and/or by increasing interactions with ECL1. To analyze this and further wean HIV-1 from human CCR5, we selected variants using CCR5(HMMH) with murine ECL1 and 2 sequences. HIV-1JRCSF mutations adaptive for CCR5(Δ18) and CCR5(HHMH) were generally maladaptive for CCR5(HMMH), whereas the converse was true for CCR5(HMMH) adaptations. The HIV-1JRCSF variant adapted to CCR5(HMMH) also weakly used intact NIH/Swiss mouse CCR5. Our results strongly suggest that HIV-1JRCSF makes functionally critical contacts with human ECL1 and that adaptation to murine ECL1 requires multiple mutations in the crown of gp120's V3 loop.

  17. Unintended targeting of Dmp1-Cre reveals a critical role for Bmpr1a signaling in the gastrointestinal mesenchyme of adult mice

    Science.gov (United States)

    Lim, Joohyun; Burclaff, Joseph; He, Guangxu; Mills, Jason C; Long, Fanxin

    2017-01-01

    Cre/loxP technology has been widely used to study cell type-specific functions of genes. Proper interpretation of such data critically depends on a clear understanding of the tissue specificity of Cre expression. The Dmp1-Cre mouse, expressing Cre from a 14-kb DNA fragment of the mouse Dmp1 gene, has become a common tool for studying gene function in osteocytes, but the presumed cell specificity is yet to be fully established. By using the Ai9 reporter line that expresses a red fluorescent protein upon Cre recombination, we find that in 2-month-old mice, Dmp1-Cre targets not only osteocytes within the bone matrix but also osteoblasts on the bone surface and preosteoblasts at the metaphyseal chondro-osseous junction. In the bone marrow, Cre activity is evident in certain stromal cells adjacent to the blood vessels, but not in adipocytes. Outside the skeleton, Dmp1-Cre marks not only the skeletal muscle fibers, certain cells in the cerebellum and the hindbrain but also gastric and intestinal mesenchymal cells that express Pdgfra. Confirming the utility of Dmp1-Cre in the gastrointestinal mesenchyme, deletion of Bmpr1a with Dmp1-Cre causes numerous large polyps along the gastrointestinal tract, consistent with prior work involving inhibition of BMP signaling. Thus, caution needs to be exercised when using Dmp1-Cre because it targets not only the osteoblast lineage at an earlier stage than previously appreciated, but also a number of non-skeletal cell types. PMID:28163952

  18. Metabolic Genetic Screens Reveal Multidimensional Regulation of Virulence Gene Expression in Listeria monocytogenes and an Aminopeptidase That Is Critical for PrfA Protein Activation.

    Science.gov (United States)

    Friedman, Sivan; Linsky, Marika; Lobel, Lior; Rabinovich, Lev; Sigal, Nadejda; Herskovits, Anat A

    2017-06-01

    Listeria monocytogenes is an environmental saprophyte and intracellular bacterial pathogen. Upon invading mammalian cells, the bacterium senses abrupt changes in its metabolic environment, which are rapidly transduced to regulation of virulence gene expression. To explore the relationship between L. monocytogenes metabolism and virulence, we monitored virulence gene expression dynamics across a library of genetic mutants grown under two metabolic conditions known to activate the virulent state: charcoal-treated rich medium containing glucose-1-phosphate and minimal defined medium containing limiting concentrations of branched-chain amino acids (BCAAs). We identified over 100 distinct mutants that exhibit aberrant virulence gene expression profiles, the majority of which mapped to nonessential metabolic genes. Mutants displayed enhanced, decreased, and early and late virulence gene expression profiles, as well as persistent levels, demonstrating a high plasticity in virulence gene regulation. Among the mutants, one was noteworthy for its particularly low virulence gene expression level and mapped to an X-prolyl aminopeptidase (PepP). We show that this peptidase plays a role in posttranslational activation of the major virulence regulator, PrfA. Specifically, PepP mediates recruitment of PrfA to the cytoplasmic membrane, a step identified as critical for PrfA protein activation. This study establishes a novel step in the complex mechanism of PrfA activation and further highlights the cross regulation of metabolism and virulence. Copyright © 2017 American Society for Microbiology.

  19. Successful bone marrow transplantation reveals the lack of endothelial progenitor cells mobilization in a patient with critical limb ischemia: a case report.

    Science.gov (United States)

    Cobellis, G; Botti, C; Taddeo, A; Silvestroni, A; Lillo, S; Da Ponte, A; Villa, M L; Sica, V; Della Bella, S

    2010-09-01

    Restoring blood flow to ischemic tissue is a prerequisite for treatment of ischemic diseases. Cell-based therapy based on bone marrow transplantation is a promising option for patients with critical limb ischemia (CLI). The efficacy of cell therapies to augment neovascularization seems to involve endothelial progenitor cells (EPCs); however, the mechanisms underlying the efficacy have not been fully elucidated. Herein we have described the case of a young patient with severe CLI, who experienced a 24-month beneficial clinical response to autologous bone marrow transplantation. The exceptional amelioration enabled him to perform standardized maximal treadmill exercise test that demonstrated lack of exercise-induced EPC mobilization, despite adequate stromal-derived factor 1 and vascular endothelial growth factor responses. Therefore, tissue ischemia is not sufficient to promote the recruitment of EPCs that have been demonstrated to be involved in the recovery from ischemia. The local implantation of marrow-derived elements may provide cells and/or trophic factors, which have the capacity to augment angiogenesis, opening new approaches to the etiopathogenesis of the disease. 2010 Elsevier Inc. All rights reserved.

  20. Studies of mice with cyclic AMP-dependent protein kinase (PKA) defects reveal the critical role of PKA's catalytic subunits in anxiety

    Science.gov (United States)

    Briassoulis, George; Keil, Margaret F.; Naved, Bilal; Liu, Sophie; Starost, Matthew F.; Nesterova, Maria; Gokarn, Nirmal; Batistatos, Anna; Wu, T. John; Stratakis, Constantine A.

    2016-01-01

    Cyclic adenosine mono-phosphate-dependent protein kinase (PKA) is critically involved in the regulation of behavioral responses. Previous studies showed that PKA's main regulatory subunit, R1α, is involved in anxiety-like behaviors. The purpose of this study was to determine how the catalytic subunit, Cα, might affect R1α's function and determine its effects on anxiety-related behaviors. The marble bury (MB) and elevated plus maze (EPM) tests were used to assess anxiety-like behavior and the hotplate test to assess nociception in wild type (WT) mouse, a Prkar1a heterozygote (Prkar1a+/-) mouse with haploinsufficiency for the regulatory subunit (R1α), a Prkaca heterozygote (Prkaca+/-) mouse with haploinsufficiency for the catalytic subunit (Cα), and a double heterozygote mouse (Prkar1a+/-/Prkaca+/-) with haploinsufficiency for both R1α and Cα. We then examined specific brain nuclei involved in anxiety. Results of MB test showed a genotype effect, with increased anxiety-like behavior in Prkar1a+/- and Prkar1a+/-/Prkaca+/- compared to WT mice. In the EPM, Prkar1a+/- spent significantly less time in the open arms, while Prkaca+/- and Prkar1a+/-/Prkaca+/- mice displayed less exploratory behavior compared to WT mice. The loss of one Prkar1a allele was associated with a significant increase in PKA activity in the basolateral (BLA) and central (CeA) amygdala and ventromedial hypothalamus (VMH) in both Prkar1a+/- and Prkar1a+/- /Prkaca+/- mice. Alterations of PKA activity induced by haploinsufficiency of its main regulatory or most important catalytic subunits result in anxiety-like behaviors. The BLA, CeA, and VMH are implicated in mediating these PKA effects in brain. PMID:26992826

  1. A Systems Approach Reveals MAVS Signaling in Myeloid Cells as Critical for Resistance to Ebola Virus in Murine Models of Infection

    Directory of Open Access Journals (Sweden)

    Mukta Dutta

    2017-01-01

    Full Text Available The unprecedented 2013–2016 outbreak of Ebola virus (EBOV resulted in over 11,300 human deaths. Host resistance to RNA viruses requires RIG-I-like receptor (RLR signaling through the adaptor protein, mitochondrial antiviral signaling protein (MAVS, but the role of RLR-MAVS in orchestrating anti-EBOV responses in vivo is not known. Here we apply a systems approach to MAVS−/− mice infected with either wild-type or mouse-adapted EBOV. MAVS controlled EBOV replication through the expression of IFNα, regulation of inflammatory responses in the spleen, and prevention of cell death in the liver, with macrophages implicated as a major cell type influencing host resistance. A dominant role for RLR signaling in macrophages was confirmed following conditional MAVS deletion in LysM+ myeloid cells. These findings reveal tissue-specific MAVS-dependent transcriptional pathways associated with resistance to EBOV, and they demonstrate that EBOV adaptation to cause disease in mice involves changes in two distinct events, RLR-MAVS antagonism and suppression of RLR-independent IFN-I responses.

  2. A Boolean Function for Neural Induction Reveals a Critical Role of Direct Intercellular Interactions in Patterning the Ectoderm of the Ascidian Embryo.

    Science.gov (United States)

    Ohta, Naoyuki; Waki, Kana; Mochizuki, Atsushi; Satou, Yutaka

    2015-12-01

    A complex system of multiple signaling molecules often produce differential gene expression patterns in animal embryos. In the ascidian embryo, four signaling ligands, Ephrin-A.d (Efna.d), Fgf9/16/20, Admp, and Gdf1/3-r, coordinately induce Otx expression in the neural lineage at the 32-cell stage. However, it has not been determined whether differential inputs of all of these signaling pathways are really necessary. It is possible that differential activation of one of these signaling pathways is sufficient and the remaining signaling pathways are activated in all cells at similar levels. To address this question, we developed a parameter-free method for determining a Boolean function for Otx expression in the present study. We treated activities of signaling pathways as Boolean values, and we also took all possible patterns of signaling gradients into consideration. We successfully determined a Boolean function that explains Otx expression in the animal hemisphere of wild-type and morphant embryos at the 32-cell stage. This Boolean function was not inconsistent with three sensing patterns, which represented whether or not individual cells received sufficient amounts of the signaling molecules. These sensing patterns all indicated that differential expression of Otx in the neural lineage is primarily determined by Efna.d, but not by differential inputs of Fgf9/16/20, Admp, and Gdf1/3-r signaling. To confirm this hypothesis experimentally, we simultaneously knocked-down Admp, Gdf1/3-r, and Fgf9/16/20, and treated this triple morphant with recombinant bFGF and BMP4 proteins, which mimic Fgf9/16/20 and Admp/Gdf1/3-r activity, respectively. Although no differential inputs of Admp, Gdf1/3-r and Fgf9/16/20 signaling were expected under this experimental condition, Otx was expressed specifically in the neural lineage. Thus, direct cell-cell interactions through Efna.d play a critical role in patterning the ectoderm of the early ascidian embryo.

  3. QM/MM free-energy simulations of reaction in Serratia marcescens Chitinase B reveal the protonation state of Asp142 and the critical role of Tyr214.

    Science.gov (United States)

    Jitonnom, Jitrayut; Limb, Michael A L; Mulholland, Adrian J

    2014-05-01

    Serratia marcescens Chitinase B (ChiB), belonging to the glycosidase family 18 (GH18), catalyzes the hydrolysis of β-1,4-glycosidic bond, with retention of configuration, via an unusual substrate-assisted mechanism, in which the substrate itself acts as an intramolecular nucleophile. Here, both elementary steps (glycosylation and deglycosylation) of the ChiB-catalyzed reaction are investigated by means of combined quantum mechanics/molecular mechanics (QM/MM) umbrella sampling molecular dynamics (MD) simulations at the SCC-DFTB/CHARMM22 level of theory. We examine the influence of the Asp142 protonation state on the reaction and the role that this residue performs in the reaction. Our simulations show that reaction with a neutral Asp142 is preferred and demonstrate that this residue provides electrostatic stabilization of the oxazolinium ion intermediate formed in the reaction. Insight into the conformational itinerary ((1,4)B↔(4)H5↔(4)C1) adopted by the substrate (bound in subsite -1) along the preferred reaction pathway is also provided by the simulations. The relative energies of the stationary points found along the reaction pathway calculated with SCC-DFTB and B3LYP were compared. The results suggest that SCC-DFTB is an accurate method for estimating the relative barriers for both steps of the reaction; however, it was found to overestimate the relative energy of an intermediate formed in the reaction when compared with the higher level of theory. Glycosylation is suggested to be a rate-determining step in the reaction with calculated overall reaction free-energy barrier of 20.5 kcal/mol, in a reasonable agreement with the 16.1 kcal/mol barrier derived from the experiment. The role of Tyr214 in catalysis was also investigated with the results, indicating that the residue plays a critical role in the deglycosylation step of the reaction. Simulations of the enzyme-product complex were also performed with an unbinding event suggested to have been observed

  4. Bacterial Hypoxic Responses Revealed as Critical Determinants of the Host-Pathogen Outcome by TnSeq Analysis of Staphylococcus aureus Invasive Infection.

    Directory of Open Access Journals (Sweden)

    Aimee D Wilde

    2015-12-01

    Full Text Available Staphylococcus aureus is capable of infecting nearly every organ in the human body. In order to infiltrate and thrive in such diverse host tissues, staphylococci must possess remarkable flexibility in both metabolic and virulence programs. To investigate the genetic requirements for bacterial survival during invasive infection, we performed a transposon sequencing (TnSeq analysis of S. aureus during experimental osteomyelitis. TnSeq identified 65 genes essential for staphylococcal survival in infected bone and an additional 148 mutants with compromised fitness in vivo. Among the loci essential for in vivo survival was SrrAB, a staphylococcal two-component system previously reported to coordinate hypoxic and nitrosative stress responses in vitro. Healthy bone is intrinsically hypoxic, and intravital oxygen monitoring revealed further decreases in skeletal oxygen concentrations upon S. aureus infection. The fitness of an srrAB mutant during osteomyelitis was significantly increased by depletion of neutrophils, suggesting that neutrophils impose hypoxic and/or nitrosative stresses on invading bacteria. To more globally evaluate staphylococcal responses to changing oxygenation, we examined quorum sensing and virulence factor production in staphylococci grown under aerobic or hypoxic conditions. Hypoxic growth resulted in a profound increase in quorum sensing-dependent toxin production, and a concomitant increase in cytotoxicity toward mammalian cells. Moreover, aerobic growth limited quorum sensing and cytotoxicity in an SrrAB-dependent manner, suggesting a mechanism by which S. aureus modulates quorum sensing and toxin production in response to environmental oxygenation. Collectively, our results demonstrate that bacterial hypoxic responses are key determinants of the staphylococcal-host interaction.

  5. Structural modeling and mutational analysis of yeast eukaryotic translation initiation factor 5A reveal new critical residues and reinforce its involvement in protein synthesis

    Science.gov (United States)

    Dias, Camila A. O.; Cano, Veridiana S. P.; Rangel, Suzana M.; Apponi, Luciano H.; Frigieri, Mariana C.; Muniz, João R. C.; Garcia, Wanius; Park, Myung H.; Garratt, Richard C.; Zanelli, Cleslei F.; Valentini, Sandro R.

    2017-01-01

    Eukaryotic translation initiation factor 5A (eIF5A) is a protein that is highly conserved and essential for cell viability. This factor is the only protein known to contain the unique and essential amino acid residue hypusine. This work focused on the structural and functional characterization of Saccharomyces cerevisiae eIF5A. The tertiary structure of yeast eIF5A was modeled based on the structure of its Leishmania mexicana homologue and this model was used to predict the structural localization of new site-directed and randomly generated mutations. Most of the 40 new mutants exhibited phenotypes that resulted from eIF-5A protein-folding defects. Our data provided evidence that the C-terminal α-helix present in yeast eIF5A is an essential structural element, whereas the eIF5A N-terminal 10 amino acid extension not present in archaeal eIF5A homologs, is not. Moreover, the mutants containing substitutions at or in the vicinity of the hypusine modification site displayed nonviable or temperature-sensitive phenotypes and were defective in hypusine modification. Interestingly, two of the temperature-sensitive strains produced stable mutant eIF5A proteins – eIF5AK56A and eIF5AQ22H,L93F – and showed defects in protein synthesis at the restrictive temperature. Our data revealed important structural features of eIF5A that are required for its vital role in cell viability and underscored an essential function of eIF5A in the translation step of gene expression. PMID:18341589

  6. Molecular dynamics simulation of human serum paraoxonase 1 in DPPC bilayer reveals a critical role of transmembrane helix H1 for HDL association.

    Science.gov (United States)

    Patra, Mahesh Chandra; Rath, Surya Narayan; Pradhan, Sukanta Kumar; Maharana, Jitendra; De, Sachinandan

    2014-01-01

    Serum paraoxonase 1 (PON1) is a high-density lipoprotein (HDL)-bound mammalian enzyme exhibiting antiatherosclerotic activity. Despite years of research, an accurate model for the binding interaction between PON1 and HDL has not been established. However, it is reported that anchoring of PON1 to HDL is mainly governed by an N-terminal alpha helix H1 and another short helix H2. Here, we studied the molecular association of full-length human PON1 (huPON1) with a HDL-mimetic dipalmitoylphosphatidylcholine (DPPC) bilayer using homology modeling and molecular dynamics simulations. Our results indicate that H1 is the highly dynamic part of huPON1, showing clockwise rotation of up to 30° within the DPPC bilayer. However, without phospholipid molecules, H1 experiences helical distortions, illustrating an incompatible HDL-anchoring conformation. Snorkeling interactions of K3, R18, and R27 together with aromatic locks formed by Y187, Y190, W194, and W202 are highly essential for anchoring of huPON1 to HDL's surface. Molecular mechanics/Poisson-Boltzmann solvent-accessible surface area (MM/PBSA) binding free energy calculation revealed that H1 displays greater binding affinity towards lipid molecules compared with H2 and H3, suggesting that H1 is the most probable HDL-binding domain of PON1. Binding free energy decomposition showed that K3, R18, and R27 interact with polar headgroups of DPPC membrane through electrostatic interaction. Moreover, Y187, Y190, W194, and W202 interact with DPPC lipids mainly through van der Waals interaction. Taken together, these results show that the transmembrane helix H1 along with the interfacial positively charged and aromatic resides were crucial for PON1's association with HDL particle. The current study will be useful towards understanding the antiatherosclerotic and bioscavenging properties of this promiscuous enzyme.

  7. A Phase I Double Blind, Placebo-Controlled, Randomized Study of the Safety and Immunogenicity of an Adjuvanted HIV-1 Gag-Pol-Nef Fusion Protein and Adenovirus 35 Gag-RT-Int-Nef Vaccine in Healthy HIV-Uninfected African Adults

    Science.gov (United States)

    Omosa-Manyonyi, Gloria; Mpendo, Juliet; Ruzagira, Eugene; Kilembe, William; Chomba, Elwyn; Roman, François; Bourguignon, Patricia; Koutsoukos, Marguerite; Collard, Alix; Voss, Gerald; Laufer, Dagna; Stevens, Gwynn; Hayes, Peter; Clark, Lorna; Cormier, Emmanuel; Dally, Len; Barin, Burc; Ackland, Jim; Syvertsen, Kristen; Zachariah, Devika; Anas, Kamaal; Sayeed, Eddy; Lombardo, Angela; Gilmour, Jill; Cox, Josephine; Fast, Patricia; Priddy, Frances

    2015-01-01

    Background Sequential prime-boost or co-administration of HIV vaccine candidates based on an adjuvanted clade B p24, RT, Nef, p17 fusion protein (F4/AS01) plus a non-replicating adenovirus 35 expressing clade A Gag, RT, Int and Nef (Ad35-GRIN) may lead to a unique immune profile, inducing both strong T-cell and antibody responses. Methods In a phase 1, double-blind, placebo-controlled trial, 146 healthy adult volunteers were randomized to one of four regimens: heterologous prime-boost with two doses of F4/AS01E or F4/AS01B followed by Ad35-GRIN; Ad35-GRIN followed by two doses of F4/AS01B; or three co-administrations of Ad35-GRIN and F4/AS01B. T cell and antibody responses were measured. Results The vaccines were generally well-tolerated, and did not cause serious adverse events. The response rate, by IFN-γ ELISPOT, was greater when Ad35-GRIN was the priming vaccine and in the co-administration groups. F4/AS01 induced CD4+ T-cells expressing primarily CD40L and IL2 +/- TNF-α, while Ad35-GRIN induced predominantly CD8+ T-cells expressing IFN-γ +/- IL2 or TNF-α. Viral inhibition was induced after Ad35-GRIN vaccination, regardless of the regimen. Strong F4-specific antibody responses were induced. Immune responses persisted at least a year after the last vaccination. The complementary response profiles, characteristic of each vaccine, were both expressed after co-administration. Conclusion Co-administration of an adjuvanted protein and an adenovirus vector showed an acceptable safety and reactogenicity profile and resulted in strong, multifunctional and complementary HIV-specific immune responses. Trial Registration ClinicalTrials.gov NCT01264445 PMID:25961283

  8. HIV-specific CD8~+ T cell responses to HXB2 Gag and Nef peptide pools in Chinese HIV/AIDS patients

    Institute of Scientific and Technical Information of China (English)

    ZHANG HongWei; QIU ZhiFeng; JIAO Yang; WANG AiXia; LI TaiSheng

    2009-01-01

    HXB2 is primarily used as a template strain in developing HIV vaccines in Europe and the US. However, it is not yet known whether the strain can induce strong HIV-specific CD8~+ T cell responses in Chinese HIV/AIDS patients. In the present study, two groups of subjects were investigated: 9 AIDS patients and 7 long-term nonprogressors (LTNPs). HIV-specific CD8~+ T cell responses were examined in all patients through the ELISPOT assay. CD4~+ T cell counts, CD8~+ T cell counts, viral load and HIV subtype of each patient were also measured. Thailand B virus strain was identified among all the patients. The breadth and magnitude of HIV-specific CD8~+ T cell responses in the LTNPs group are greater than those in the AIDS group (P<0.01). There is a positive correlation between magnitude of HIV-specific CD8~+ T cell responses and CD4~+ T cells, and a negative correlation between HIV-specific CD8~+ T cell responses and mean viral load. In summary, the HIV-specific CD8~+ T cell responses to the HXB2 Gag and Nef peptide pools are considerable in Chinese HIV/AIDS patients infected with Thailand B virus strain. HIV-1 vaccines based on HXB2 strain that can induce extensive immunity may be helpful for Chinese.

  9. Engineering and Validation of a Vector for Concomitant Expression of Rare Transfer RNA (tRNA) and HIV-1 nef Genes in Escherichia coli.

    Science.gov (United States)

    Mualif, Siti Aisyah; Teow, Sin-Yeang; Omar, Tasyriq Che; Chew, Yik Wei; Yusoff, Narazah Mohd; Ali, Syed A

    2015-01-01

    Relative ease in handling and manipulation of Escherichia coli strains make them primary candidate to express proteins heterologously. Overexpression of heterologous genes that contain codons infrequently used by E. coli is related with difficulties such as mRNA instability, early termination of transcription and/or translation, deletions and/or misincorporation, and cell growth inhibition. These codon bias -associated problems are addressed by co-expressing ColE1-compatible, rare tRNA expressing helper plasmids. However, this approach has inadequacies, which we have addressed by engineering an expression vector that concomitantly expresses the heterologous protein of interest, and rare tRNA genes in E. coli. The expression vector contains three (argU, ileY, leuW) rare tRNA genes and a useful multiple cloning site for easy in-frame cloning. To maintain the overall size of the parental plasmid vector, the rare tRNA genes replaced the non-essential DNA segments in the vector. The cloned gene is expressed under the control of T7 promoter and resulting recombinant protein has a C-terminal 6His tag for IMAC-mediated purification. We have evaluated the usefulness of this expression vector by expressing three HIV-1 genes namely HIV-1 p27 (nef), HIV-1 p24 (ca), and HIV-1 vif in NiCo21(DE3) E.coli and demonstrated the advantages of using expression vector that concomitantly expresses rare tRNA and heterologous genes.

  10. HIV-specific CD8~+ T cell responses to HXB2 Gag and Nef peptide pools in Chinese HIV/AIDS patients

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    HXB2 is primarily used as a template strain in developing HIV vaccines in Europe and the US. However,it is not yet known whether the strain can induce strong HIV-specific CD8+ T cell responses in Chinese HIV/AIDS patients. In the present study,two groups of subjects were investigated:9 AIDS patients and 7 long-term nonprogressors (LTNPs). HIV-specific CD8+ T cell responses were examined in all patients through the ELISPOT assay. CD4+ T cell counts,CD8+ T cell counts,viral load and HIV subtype of each patient were also measured. Thailand B virus strain was identified among all the patients. The breadth and magnitude of HIV-specific CD8+ T cell responses in the LTNPs group are greater than those in the AIDS group (P<0.01). There is a positive correlation between magnitude of HIV-specific CD8+ T cell responses and CD4+ T cells,and a negative correlation between HIV-specific CD8+ T cell responses and mean viral load. In summary,the HIV-specific CD8+ T cell responses to the HXB2 Gag and Nef peptide pools are considerable in Chinese HIV/AIDS patients infected with Thailand B virus strain. HIV-1 vaccines based on HXB2 strain that can induce extensive immunity may be helpful for Chinese.

  11. Increased Cardiovascular Disease Risk In The HIV-Positive Population on ART: Potential Role of HIV-Nef and Tat

    OpenAIRE

    Wang, Ting; Yi, Ru; Green, Linden Ann; Chelvanambi, Sarvesh; Seimetz, Michael; Clauss, Matthias

    2015-01-01

    With effective antiretroviral therapy (ART), many HIV-infected people die of diseases other than acquired immune deficiency syndrome (AIDS). In particular, coronary artery disease has emerged as one of most critical complications of HIV infection and a major cause of morbidity and mortality. Although reportedly antiretroviral combination therapy itself may accelerate atherosclerosis by enhancing dyslipidemia, most recent epidemiological studies support the notion that HIV infection itself con...

  12. CriticalEd

    DEFF Research Database (Denmark)

    Kjellberg, Caspar Mølholt; Meredith, David

    2014-01-01

    The best text method is commonly applied among music scholars engaged in producing critical editions. In this method, a comment list is compiled, consisting of variant readings and editorial emendations. This list is maintained by inserting the comments into a document as the changes are made....... Since the comments are not input sequentially, with regard to position, but in arbitrary order, this list must be sorted by copy/pasting the rows into place—an error-prone and time-consuming process. Scholars who produce critical editions typically use off-the-shelf music notation software...... such as Sibelius or Finale. It was hypothesized that it would be possible to develop a Sibelius plug-in, written in Manuscript 6, that would improve the critical editing work flow, but it was found that the capabilities of this scripting language were insufficient. Instead, a 3-part system was designed and built...

  13. Location and dynamics of the immunodominant CD8 T cell response to SIVΔnef immunization and SIVmac251 vaginal challenge.

    Directory of Open Access Journals (Sweden)

    Arun K Sasikala-Appukuttan

    Full Text Available Live-attenuated SIV vaccines (LAVs have been the most effective to date in preventing or partially controlling infection by wild-type SIV in non-human primate models of HIV-1 transmission to women acting by mechanisms of protection that are not well understood. To gain insights into mechanisms of protection by LAVs that could aid development of effective vaccines to prevent HIV-1 transmission to women, we used in situ tetramer staining to determine whether increased densities or changes in the local distribution of SIV-specific CD8 T cells correlated with the maturation of SIVΔnef vaccine-induced protection prior to and after intra-vaginal challenge with wild-type SIVmac251. We evaluated the immunodominant Mamu-A1*001:01/Gag (CM9 and Mamu-A1*001:01/Tat (SL8 epitope response in genital and lymphoid tissues, and found that tetramer+ cells were present at all time points examined. In the cervical vaginal tissues, most tetramer+ cells were distributed diffusely throughout the lamina propria or co-localized with other CD8 T cells within lymphoid aggregates. The distribution and densities of the tetramer+ cells at the portal of entry did not correlate with the maturation of protection or change after challenge. Given these findings, we discuss the possibility that changes in other aspects of the immune system, including the quality of the resident population of virus-specific effector CD8 T cells could contribute to maturation of protection, as well as the potential for vaccine strategies that further increase the size and quality of this effector population to prevent HIV-1 transmission.

  14. Engineering and Validation of a Vector for Concomitant Expression of Rare Transfer RNA (tRNA and HIV-1 nef Genes in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Siti Aisyah Mualif

    Full Text Available Relative ease in handling and manipulation of Escherichia coli strains make them primary candidate to express proteins heterologously. Overexpression of heterologous genes that contain codons infrequently used by E. coli is related with difficulties such as mRNA instability, early termination of transcription and/or translation, deletions and/or misincorporation, and cell growth inhibition. These codon bias -associated problems are addressed by co-expressing ColE1-compatible, rare tRNA expressing helper plasmids. However, this approach has inadequacies, which we have addressed by engineering an expression vector that concomitantly expresses the heterologous protein of interest, and rare tRNA genes in E. coli. The expression vector contains three (argU, ileY, leuW rare tRNA genes and a useful multiple cloning site for easy in-frame cloning. To maintain the overall size of the parental plasmid vector, the rare tRNA genes replaced the non-essential DNA segments in the vector. The cloned gene is expressed under the control of T7 promoter and resulting recombinant protein has a C-terminal 6His tag for IMAC-mediated purification. We have evaluated the usefulness of this expression vector by expressing three HIV-1 genes namely HIV-1 p27 (nef, HIV-1 p24 (ca, and HIV-1 vif in NiCo21(DE3 E.coli and demonstrated the advantages of using expression vector that concomitantly expresses rare tRNA and heterologous genes.

  15. Increased cardiovascular disease risk in the HIV-positive population on ART: potential role of HIV-Nef and Tat.

    Science.gov (United States)

    Wang, Ting; Yi, Ru; Green, Linden Ann; Chelvanambi, Sarvesh; Seimetz, Michael; Clauss, Matthias

    2015-01-01

    With effective antiretroviral therapy (ART), many HIV-infected people die of diseases other than acquired immune deficiency syndrome (AIDS). In particular, coronary artery disease has emerged as one of most critical complications of HIV infection and a major cause of morbidity and mortality. Although reportedly antiretroviral combination therapy itself may accelerate atherosclerosis by enhancing dyslipidemia, most recent epidemiological studies support the notion that HIV infection itself contributes to cardiovascular disease. However, it is still a mystery how the virus can contribute to cardiovascular disease development even while suppressed by ARTs. This review discusses the current understanding of interactions between HIV infection and cardiovascular diseases in both clinical and experimental studies with special focus on those viral proteins that are still produced by HIV. This will help infectious disease/vascular biology experts to gain insights into the pathophysiological mechanisms of HIV-associated cardiovascular disease and new trends to treat and prevent cardiovascular disease in the HIV-infected population.

  16. Interleukin-1- and type I interferon-dependent enhanced immunogenicity of an NYVAC-HIV-1 Env-Gag-Pol-Nef vaccine vector with dual deletions of type I and type II interferon-binding proteins.

    Science.gov (United States)

    Delaloye, Julie; Filali-Mouhim, Abdelali; Cameron, Mark J; Haddad, Elias K; Harari, Alexandre; Goulet, Jean-Pierre; Gomez, Carmen E; Perdiguero, Beatriz; Esteban, Mariano; Pantaleo, Giuseppe; Roger, Thierry; Sékaly, Rafick-Pierre; Calandra, Thierry

    2015-04-01

    NYVAC, a highly attenuated, replication-restricted poxvirus, is a safe and immunogenic vaccine vector. Deletion of immune evasion genes from the poxvirus genome is an attractive strategy for improving the immunogenic properties of poxviruses. Using systems biology approaches, we describe herein the enhanced immunological profile of NYVAC vectors expressing the HIV-1 clade C env, gag, pol, and nef genes (NYVAC-C) with single or double deletions of genes encoding type I (ΔB19R) or type II (ΔB8R) interferon (IFN)-binding proteins. Transcriptomic analyses of human monocytes infected with NYVAC-C, NYVAC-C with the B19R deletion (NYVAC-C-ΔB19R), or NYVAC-C with B8R and B19R deletions (NYVAC-C-ΔB8RB19R) revealed a concerted upregulation of innate immune pathways (IFN-stimulated genes [ISGs]) of increasing magnitude with NYVAC-C-ΔB19R and NYVAC-C-ΔB8RB19R than with NYVAC-C. Deletion of B8R and B19R resulted in an enhanced activation of IRF3, IRF7, and STAT1 and the robust production of type I IFNs and of ISGs, whose expression was inhibited by anti-type I IFN antibodies. Interestingly, NYVAC-C-ΔB8RB19R induced the production of much higher levels of proinflammatory cytokines (tumor necrosis factor [TNF], interleukin-6 [IL-6], and IL-8) than NYVAC-C or NYVAC-C-ΔB19R as well as a strong inflammasome response (caspase-1 and IL-1β) in infected monocytes. Top network analyses showed that this broad response mediated by the deletion of B8R and B19R was organized around two upregulated gene expression nodes (TNF and IRF7). Consistent with these findings, monocytes infected with NYVAC-C-ΔB8RB19R induced a stronger type I IFN-dependent and IL-1-dependent allogeneic CD4(+) T cell response than monocytes infected with NYVAC-C or NYVAC-C-ΔB19R. Dual deletion of type I and type II IFN immune evasion genes in NYVAC markedly enhanced its immunogenic properties via its induction of the increased expression of type I IFNs and IL-1β and make it an attractive candidate HIV

  17. Immunogenicity of seven new recombinant yellow fever viruses 17D expressing fragments of SIVmac239 Gag, Nef, and Vif in Indian rhesus macaques.

    Directory of Open Access Journals (Sweden)

    Mauricio A Martins

    Full Text Available An effective vaccine remains the best solution to stop the spread of human immunodeficiency virus (HIV. Cellular immune responses have been repeatedly associated with control of viral replication and thus may be an important element of the immune response that must be evoked by an efficacious vaccine. Recombinant viral vectors can induce potent T-cell responses. Although several viral vectors have been developed to deliver HIV genes, only a few have been advanced for clinical trials. The live-attenuated yellow fever vaccine virus 17D (YF17D has many properties that make it an attractive vector for AIDS vaccine regimens. YF17D is well tolerated in humans and vaccination induces robust T-cell responses that persist for years. Additionally, methods to manipulate the YF17D genome have been established, enabling the generation of recombinant (rYF17D vectors carrying genes from unrelated pathogens. Here, we report the generation of seven new rYF17D viruses expressing fragments of simian immunodeficiency virus (SIVmac239 Gag, Nef, and Vif. Studies in Indian rhesus macaques demonstrated that these live-attenuated vectors replicated in vivo, but only elicited low levels of SIV-specific cellular responses. Boosting with recombinant Adenovirus type-5 (rAd5 vectors resulted in robust expansion of SIV-specific CD8(+ T-cell responses, particularly those targeting Vif. Priming with rYF17D also increased the frequency of CD4(+ cellular responses in rYF17D/rAd5-immunized macaques compared to animals that received rAd5 only. The effect of the rYF17D prime on the breadth of SIV-specific T-cell responses was limited and we also found evidence that some rYF17D vectors were more effective than others at priming SIV-specific T-cell responses. Together, our data suggest that YF17D - a clinically relevant vaccine vector - can be used to prime AIDS virus-specific T-cell responses in heterologous prime boost regimens. However, it will be important to optimize rYF17D

  18. Thinking Critically about Critical Thinking

    Science.gov (United States)

    Mulnix, Jennifer Wilson

    2012-01-01

    As a philosophy professor, one of my central goals is to teach students to think critically. However, one difficulty with determining whether critical thinking can be taught, or even measured, is that there is widespread disagreement over what critical thinking actually is. Here, I reflect on several conceptions of critical thinking, subjecting…

  19. d-f hybridization and quantum criticality in weakly-itinerant ferromagnets.

    Science.gov (United States)

    Silva Neto, M B; Castro Neto, A H; Kim, J S; Stewart, G R

    2013-01-16

    We investigate the unusual magnetic, thermodynamic and transport properties of nearly-critical, weakly-itinerant ferromagnets with the general formula UTX, where T=Rh, Co and X=Ge, Si. As a unique feature of these systems, we show how changes in the V(df) hybridization, which controls their proximity to a ferromagnetic instability, determine the evolution of the ground state magnetization, M(0), the Curie temperature, T(C), the density of states at the Fermi level, N(E(F)), the T(2) resistivity coefficient, A, and the specific heat coefficient, γ. The universal aspect of our findings comes from the dependence on only two parameters: the transition metal T(d) bandwidth, W(d), and the distance between the T(d) and U(f) band centers, C(T(d)) - C(U(f)). We discuss our results in connection to data for URh(1-x)Co(x)Ge.

  20. Critical Care

    Science.gov (United States)

    Critical care helps people with life-threatening injuries and illnesses. It might treat problems such as complications ... a team of specially-trained health care providers. Critical care usually takes place in an intensive care ...

  1. Archetypal Criticism.

    Science.gov (United States)

    Chesebro, James W.; And Others

    1990-01-01

    Argues that archetypal criticism is a useful way of examining universal, historical, and cross-cultural symbols in classrooms. Identifies essential features of an archetype; outlines operational and critical procedures; illustrates archetypal criticism as applied to the cross as a symbol; and provides a synoptic placement for archetypal criticism…

  2. How Critical Is Critical Thinking?

    Science.gov (United States)

    Shaw, Ryan D.

    2014-01-01

    Recent educational discourse is full of references to the value of critical thinking as a 21st-century skill. In music education, critical thinking has been discussed in relation to problem solving and music listening, and some researchers suggest that training in critical thinking can improve students' responses to music. But what exactly is…

  3. Critical Jostling

    Directory of Open Access Journals (Sweden)

    Pippin Barr

    2016-11-01

    Full Text Available Games can serve a critical function in many different ways, from serious games about real world subjects to self-reflexive commentaries on the nature of games themselves. In this essay we discuss critical possibilities stemming from the area of critical design, and more specifically Carl DiSalvo’s adversarial design and its concept of reconfiguring the remainder. To illustrate such an approach, we present the design and outcomes of two games, Jostle Bastard and Jostle Parent. We show how the games specifically engage with two previous games, Hotline Miami and Octodad: Dadliest Catch, reconfiguring elements of those games to create interactive critical experiences and extensions of the source material. Through the presentation of specific design concerns and decisions, we provide a grounded illustration of a particular critical function of videogames and hope to highlight this form as another valuable approach in the larger area of videogame criticism.

  4. Criticality Model

    Energy Technology Data Exchange (ETDEWEB)

    A. Alsaed

    2004-09-14

    The ''Disposal Criticality Analysis Methodology Topical Report'' (YMP 2003) presents the methodology for evaluating potential criticality situations in the monitored geologic repository. As stated in the referenced Topical Report, the detailed methodology for performing the disposal criticality analyses will be documented in model reports. Many of the models developed in support of the Topical Report differ from the definition of models as given in the Office of Civilian Radioactive Waste Management procedure AP-SIII.10Q, ''Models'', in that they are procedural, rather than mathematical. These model reports document the detailed methodology necessary to implement the approach presented in the Disposal Criticality Analysis Methodology Topical Report and provide calculations utilizing the methodology. Thus, the governing procedure for this type of report is AP-3.12Q, ''Design Calculations and Analyses''. The ''Criticality Model'' is of this latter type, providing a process evaluating the criticality potential of in-package and external configurations. The purpose of this analysis is to layout the process for calculating the criticality potential for various in-package and external configurations and to calculate lower-bound tolerance limit (LBTL) values and determine range of applicability (ROA) parameters. The LBTL calculations and the ROA determinations are performed using selected benchmark experiments that are applicable to various waste forms and various in-package and external configurations. The waste forms considered in this calculation are pressurized water reactor (PWR), boiling water reactor (BWR), Fast Flux Test Facility (FFTF), Training Research Isotope General Atomic (TRIGA), Enrico Fermi, Shippingport pressurized water reactor, Shippingport light water breeder reactor (LWBR), N-Reactor, Melt and Dilute, and Fort Saint Vrain Reactor spent nuclear fuel (SNF). The scope of

  5. Critical Muralism

    Science.gov (United States)

    Rosette, Arturo

    2009-01-01

    This study focuses on the development and practices of Critical Muralists--community-educator-artist-leader-activists--and situates these specifically in relation to the Mexican mural tradition of los Tres Grandes and in relation to the history of public art more generally. The study examines how Critical Muralists address artistic and…

  6. Ambiguity Revealed

    OpenAIRE

    Subir Bose; Matthew Polisson; Ludovic Renou

    2012-01-01

    We derive necessary and suffcient conditions for data sets composed of state-contingent prices and consumption to be consistent with two prominent models of decision making under ambiguity: variational preferences and smooth ambiguity. The revealed preference conditions for the maxmin expected utility and subjective expected utility models are characterized as special cases.

  7. Ambiguity revealed

    OpenAIRE

    Bayer, Ralph-C; Bose, Subir; Polisson, Matthew; Renou, Ludovic

    2013-01-01

    We derive necessary and sufficient conditions for data sets composed of state-contingent prices and consumption to be consistent with two prominent models of decision making under uncertainty: variational preferences and smooth ambiguity. The revealed preference conditions for subjective expected utility, maxmin expected utility, and multiplier preferences are characterised as special cases. We implement our tests on data from a portfolio choice experiment.

  8. Critical proximity

    Directory of Open Access Journals (Sweden)

    Simon, Jane

    2010-01-01

    Full Text Available This essay considers how written language frames visual objects. Drawing on Michel Foucault’s response to Raymond Roussel’s obsessive description, the essay proposes a model of criticism where description might press up against its objects. This critical closeness is then mapped across the conceptual art practice and art criticism of Ian Burn. Burn attends to the differences between seeing and reading, and considers the conditions which frame how we look at images, including how we look at, and through words. The essay goes on to consider Meaghan Morris’s writing on Lynn Silverman’s photographs. Both Morris and Burn offer an alternative to a parasitic model of criticism and enact a patient way of looking across and through visual landscapes.

  9. Critical Proximity

    Directory of Open Access Journals (Sweden)

    Jane Simon

    2010-09-01

    Full Text Available This essay considers how written language frames visual objects. Drawing on Michel Foucault’s response to Raymond Roussel’s obsessive description, the essay proposes a model of criticism where description might press up against its objects. This critical closeness is then mapped across the conceptual art practice and art criticism of Ian Burn. Burn attends to the differences between seeing and reading, and considers the conditions which frame how we look at images, including how we look at, and through words. The essay goes on to consider Meaghan Morris’s writing on Lynn Silverman’s photographs. Both Morris and Burn offer an alternative to a parasitic model of criticism and enact a patient way of looking across and through visual landscapes.

  10. Mathematics revealed

    CERN Document Server

    Berman, Elizabeth

    1979-01-01

    Mathematics Revealed focuses on the principles, processes, operations, and exercises in mathematics.The book first offers information on whole numbers, fractions, and decimals and percents. Discussions focus on measuring length, percent, decimals, numbers as products, addition and subtraction of fractions, mixed numbers and ratios, division of fractions, addition, subtraction, multiplication, and division. The text then examines positive and negative numbers and powers and computation. Topics include division and averages, multiplication, ratios, and measurements, scientific notation and estim

  11. MRKAd5 HIV-1 Gag/Pol/Nef vaccine-induced T-cell responses inadequately predict distance of breakthrough HIV-1 sequences to the vaccine or viral load.

    Directory of Open Access Journals (Sweden)

    Holly Janes

    Full Text Available BACKGROUND: The sieve analysis for the Step trial found evidence that breakthrough HIV-1 sequences for MRKAd5/HIV-1 Gag/Pol/Nef vaccine recipients were more divergent from the vaccine insert than placebo sequences in regions with predicted epitopes. We linked the viral sequence data with immune response and acute viral load data to explore mechanisms for and consequences of the observed sieve effect. METHODS: Ninety-one male participants (37 placebo and 54 vaccine recipients were included; viral sequences were obtained at the time of HIV-1 diagnosis. T-cell responses were measured 4 weeks post-second vaccination and at the first or second week post-diagnosis. Acute viral load was obtained at RNA-positive and antibody-negative visits. FINDINGS: Vaccine recipients had a greater magnitude of post-infection CD8+ T cell response than placebo recipients (median 1.68% vs 1.18%; p = 0·04 and greater breadth of post-infection response (median 4.5 vs 2; p = 0·06. Viral sequences for vaccine recipients were marginally more divergent from the insert than placebo sequences in regions of Nef targeted by pre-infection immune responses (p = 0·04; Pol p = 0·13; Gag p = 0·89. Magnitude and breadth of pre-infection responses did not correlate with distance of the viral sequence to the insert (p>0·50. Acute log viral load trended lower in vaccine versus placebo recipients (estimated mean 4·7 vs 5·1 but the difference was not significant (p = 0·27. Neither was acute viral load associated with distance of the viral sequence to the insert (p>0·30. INTERPRETATION: Despite evidence of anamnestic responses, the sieve effect was not well explained by available measures of T-cell immunogenicity. Sequence divergence from the vaccine was not significantly associated with acute viral load. While point estimates suggested weak vaccine suppression of viral load, the result was not significant and more viral load data would be needed to detect

  12. Higher cross-subtype IFN-γ ELISpot responses to Gag and Nef peptides in Brazilian HIV-1 subtype B- and F1- than in C-infected subjects.

    Science.gov (United States)

    Côrtes, Fernanda Heloise; Bello, Gonzalo; Vorsatz, Carla; Pilotto, José Henrique; Guimarães, Monick Lindenmeyer; Grinsztejn, Beatriz; Veloso, Valdiléa Gonçalves; Pinto, Aguinaldo Roberto; Morgado, Mariza Gonçalves

    2013-02-04

    HIV-1 diversity has been considered a huge challenge for the HIV-1 vaccine development. To overcome it, immunogens based on centralized sequences, as consensus, have been tested. In Brazil, the co-circulation of three subtypes offers a suitable scenario to test T cell cross-subtype responses to consensus sequences. Furthermore, we included peptides based on closest viral isolates (CVI) from each subtype analyzed to compare with T cell responses detected against the consensus sequences. The study included 32 subjects infected with HIV-1 subtype B (n=13),C (n=11), and F1 (n=8). Gag and Nef-specific T cell responses were evaluated by IFN-γ-ELISpot assay. Peptides based on CVI sequences were similar to consensus in both reducing genetic distance and detecting T cell responses. A high cross-subtype response between B and F1 in both regions was observed in HIV-1 subtype B and F1-infected subjects. We also found no significant difference in responses to subtype B and C consensus peptides among subtype B-infected subjects. In contrast, the magnitude of T cell responses to consensus C peptides in the Gag region was higher than to consensus B peptides among HIV-1 subtype C-infected subjects. Regarding Nef, subtype C-infected subjects showed higher values to consensus C than to consensus F1 peptides. Moreover, subtype F1-infected subjects presented lower responses to subtype C peptides than to subtype F1 and B. A similar level of responses was detected with group M based peptides in subtype B and F1 infected subjects. However, among subtype C infected subjects, this set of peptides detected lower levels of response than consensus C. Overall, the level of cross-subtype response between subtypes B and F1 was higher than between subtype C and B or C and F1. Our data suggests that the barrier of genetic diversity in HIV-1 group M for vaccine design may be dependent on the subtypes involved. Copyright © 2012 Elsevier Ltd. All rights reserved.

  13. REVEALED ALTRUISM

    OpenAIRE

    Cox, James C; Friedman, Daniel; Sadiraj, Vjollca

    2009-01-01

    This pap er develops a theory of revealed preferences over oneís own and othersímonetary payo§s. We intro duce ìmore altruistic thanî(MAT), a partial ordering over preferences, and interpret it with known parametric mo dels. We also intro duce and illustrate ìmore generous thanî (MGT), a partial ordering over opp ortunity sets. Several recent discussions of altruism fo cus on two player extensive form games of complete information in which the Örst mover (FM) cho oses a more or less gen...

  14. Persistence of the superconducting condensate far above the critical temperature of YBa2(Cu,Zn)3O(y) revealed by c-axis optical conductivity measurements for several Zn concentrations and carrier doping levels.

    Science.gov (United States)

    Uykur, Ece; Tanaka, Kiyohisa; Masui, Takahiko; Miyasaka, Shigeki; Tajima, Setsuko

    2014-03-28

    The superconductivity precursor phenomena in high temperature cuprate superconductors is studied by direct measurements of the superconducting condensate with the use of the c-axis optical conductivity of YBa2(Cu1-xZnx)3Oy for several doping levels (p) as well as for several Zn concentrations. Both the real and imaginary parts of the optical conductivity clearly show that the superconducting carriers persist up to the high temperatures Tp that is higher than the critical temperature Tc but lower than the pseudogap temperature T*. Tp increases with reducing doping level like T*, but decreases with Zn substitution unlike T*.

  15. Critical Vidders

    DEFF Research Database (Denmark)

    Svegaard, Robin Sebastian Kaszmarczyk

    2015-01-01

    This article will introduce and take a look at a specific subset of the fan created remix videos known as vids, namely those that deal with feminist based critique of media. Through examples, it will show how fans construct and present their critique, and finally broach the topic of the critical ...

  16. Critical Vidders

    DEFF Research Database (Denmark)

    Svegaard, Robin Sebastian Kaszmarczyk

    2015-01-01

    This article will introduce and take a look at a specific subset of the fan created remix videos known as vids, namely those that deal with feminist based critique of media. Through examples, it will show how fans construct and present their critique, and finally broach the topic of the critical...

  17. Quantum Criticality

    OpenAIRE

    Keimer, Bernhard; Sachdev, Subir

    2011-01-01

    This is a review of the basic theoretical ideas of quantum criticality, and of their connection to numerous experiments on correlated electron compounds. A shortened, modified, and edited version appeared in Physics Today. This arxiv version has additional citations to the literature.

  18. Revealing Rembrandt

    Directory of Open Access Journals (Sweden)

    Andrew J Parker

    2014-04-01

    Full Text Available The power and significance of artwork in shaping human cognition is self-evident. The starting point for our empirical investigations is the view that the task of neuroscience is to integrate itself with other forms of knowledge, rather than to seek to supplant them. In our recent work, we examined a particular aspect of the appreciation of artwork using present-day functional magnetic resonance imaging (fMRI. Our results emphasised the continuity between viewing artwork and other human cognitive activities. We also showed that appreciation of a particular aspect of artwork, namely authenticity, depends upon the co-ordinated activity between the brain regions involved in multiple decision making and those responsible for processing visual information. The findings about brain function probably have no specific consequences for understanding how people respond to the art of Rembrandt in comparison with their response to other artworks. However, the use of images of Rembrandt’s portraits, his most intimate and personal works, clearly had a significant impact upon our viewers, even though they have been spatially confined to the interior of an MRI scanner at the time of viewing. Neuroscientific studies of humans viewing artwork have the capacity to reveal the diversity of human cognitive responses that may be induced by external advice or context as people view artwork in a variety of frameworks and settings.

  19. Critical reading and critical thinking Critical reading and critical thinking

    Directory of Open Access Journals (Sweden)

    Loni Kreis Taglieber

    2008-04-01

    Full Text Available The purpose of this paper is to provide, for L1 and L2 reading and writing teachers, a brief overview of the literature about critical reading and higher level thinking skills. The teaching of these skills is still neglected in some language classes in Brazil, be it in L1 or in L2 classes. Thus, this paper may also serve as a resource guide for L1 and/or L2 reading and writing teachers who want to incorporate critical reading and thinking into their classes. In modern society, even in everyday life people frequently need to deal with complicated public and political issues, make decisions, and solve problems. In order to do this efficiently and effectively, citizens must be able to evaluate critically what they see, hear, and read. Also, with the huge amount of printed material available in all areas in this age of “information explosion” it is easy to feel overwhelmed. But often the information piled up on people’s desks and in their minds is of no use due to the enormous amount of it. The purpose of this paper is to provide, for L1 and L2 reading and writing teachers, a brief overview of the literature about critical reading and higher level thinking skills. The teaching of these skills is still neglected in some language classes in Brazil, be it in L1 or in L2 classes. Thus, this paper may also serve as a resource guide for L1 and/or L2 reading and writing teachers who want to incorporate critical reading and thinking into their classes. In modern society, even in everyday life people frequently need to deal with complicated public and political issues, make decisions, and solve problems. In order to do this efficiently and effectively, citizens must be able to evaluate critically what they see, hear, and read. Also, with the huge amount of printed material available in all areas in this age of “information explosion” it is easy to feel overwhelmed. But often the information piled up on people’s desks and in their minds is of

  20. Structure/Function Analysis of Recurrent Mutations in SETD2 Protein Reveals a Critical and Conserved Role for a SET Domain Residue in Maintaining Protein Stability and Histone H3 Lys-36 Trimethylation.

    Science.gov (United States)

    Hacker, Kathryn E; Fahey, Catherine C; Shinsky, Stephen A; Chiang, Yun-Chen J; DiFiore, Julia V; Jha, Deepak Kumar; Vo, Andy H; Shavit, Jordan A; Davis, Ian J; Strahl, Brian D; Rathmell, W Kimryn

    2016-09-30

    The yeast Set2 histone methyltransferase is a critical enzyme that plays a number of key roles in gene transcription and DNA repair. Recently, the human homologue, SETD2, was found to be recurrently mutated in a significant percentage of renal cell carcinomas, raising the possibility that the activity of SETD2 is tumor-suppressive. Using budding yeast and human cell line model systems, we examined the functional significance of two evolutionarily conserved residues in SETD2 that are recurrently mutated in human cancers. Whereas one of these mutations (R2510H), located in the Set2 Rpb1 interaction domain, did not result in an observable defect in SETD2 enzymatic function, a second mutation in the catalytic domain of this enzyme (R1625C) resulted in a complete loss of histone H3 Lys-36 trimethylation (H3K36me3). This mutant showed unchanged thermal stability as compared with the wild type protein but diminished binding to the histone H3 tail. Surprisingly, mutation of the conserved residue in Set2 (R195C) similarly resulted in a complete loss of H3K36me3 but did not affect dimethylated histone H3 Lys-36 (H3K36me2) or functions associated with H3K36me2 in yeast. Collectively, these data imply a critical role for Arg-1625 in maintaining the protein interaction with H3 and specific H3K36me3 function of this enzyme, which is conserved from yeast to humans. They also may provide a refined biochemical explanation for how H3K36me3 loss leads to genomic instability and cancer.

  1. Critical scattering

    Energy Technology Data Exchange (ETDEWEB)

    Stirling, W.G. [Liverpool Univ., Dep. of Physics, Liverpool (United Kingdom); Perry, S.C. [Keele Univ. (United Kingdom). Dept. of Physics

    1996-12-31

    We outline the theoretical and experimental background to neutron scattering studies of critical phenomena at magnetic and structural phase transitions. The displacive phase transition of SrTiO{sub 3} is discussed, along with examples from recent work on magnetic materials from the rare-earth (Ho, Dy) and actinide (NpAs, NpSb, USb) classes. The impact of synchrotron X-ray scattering is discussed in conclusion. (author) 13 figs., 18 refs.

  2. Analysis of the N-terminal positively charged residues of the simian immunodeficiency virus Vif reveals a critical amino acid required for the antagonism of rhesus APOBEC3D, G, and H.

    Science.gov (United States)

    Schmitt, Kimberly; Katuwal, Miki; Wang, Yaqiong; Li, Cicy; Stephens, Edward B

    2014-01-20

    Previous studies have shown that apolipoprotein B mRNA editing, enzyme catalytic, polypeptide G (APOBEC3G; hA3G) and F (APOBEC3F; hA3F) proteins interact with a nonlinear binding site located at the N-terminal region of the HIV-1 Vif protein. We have analyzed the role of 12 positively charged amino acids of the N-terminal region of the SIV Vif. Simian-human immunodeficiency viruses (SHIV) were constructed that expressed each of these amino acid substitutions. These viruses were examined for replication in the presence of rhesus macaque APOBEC3 proteins (rhA3A-rhA3H), incorporation of the different A3 proteins into virions, and replication in rhesus macaque PBMC. Similar to other studies, we found that K27 was essential for rhA3G activity and rhA3F but was not important for restriction of SHIVΔvif by rhA3A, rhA3D or rhA3H. Our results identified the arginine at position 14 of the SIV Vif as a critical residue for virus restriction by rhA3D, rhA3G and rhA3H.

  3. How Critical Is Critical Infrastructure?

    Science.gov (United States)

    2015-09-01

    to Examine Critical Issues Underlying the Planned Rebuilding at the World Trade Center Site. 178 HVS Global Hospitality Services, 2012 Manhattan...Hotel Market Overview (Mineola, NY: HVS Global Hospitality Services, 2012, http://www.hvs.com/Content/3268.pdf. 179 “Key Office Properties,” accessed...premier real estate, luxury shopping, world class hotels, destination dining, and tourism , into an area that produces cumulative consumer spending of

  4. [Critical incidents].

    Science.gov (United States)

    Scheidegger, D

    2005-03-01

    In medicine real severe mishaps are rare. On the other hand critical incidents are frequent. Anonymous critical incident reporting systems allow us to learn from these mishaps. This learning process will make our daily clinical work safer Unfortunately, before these systems can be used efficiently our professional culture has to be changed. Everyone in medicine has to admit that errors do occur to see the need for an open discussion. If we really want to learn from errors, we cannot punish the individual, who reported his or her mistake. The interest is primarily in what has happened and why it has happened and not who has committed this mistake. The cause for critical incidents in medicine is in over 80% the human factor Poor communication, work under enormous stress, conflicts and hierarchies are the main cause. This has been known for many years, therefore have already 15 years ago high-tech industries, like e.g. aviation, started to invest in special courses on team training. Medicine is a typical profession were until now only the individual performance decided about the professional career Communication, conflict management, stress management, decision making, risk management, team and team resource management were subjects that have never been taught during our preor postgraduate education. These points are the most important ones for an optimal teamwork. A multimodular course designed together with Swissair (Human Aspect Development medical, HADmedical) helps to cover, as in aviation, the soft factor and behavioural education in medicine and to prepare professionals in health care to work as a real team.

  5. Critical reading and critical thinking

    Directory of Open Access Journals (Sweden)

    Loni Kreis Taglieber

    2003-01-01

    Full Text Available The purpose of this paper is to provide, for L1 and L2 reading and writing teachers, a brief overview of the literature about critical reading and higher level thinking skills. The teaching of these skills is still neglected in some language classes in Brazil, be it in L1 or in L2 classes. Thus, this paper may also serve as a resource guide for L1 and/or L2 reading and writing teachers who want to incorporate critical reading and thinking into their classes. In modern society, even in everyday life people frequently need to deal with complicated public and political issues, make decisions, and solve problems. In order to do this efficiently and effectively, citizens must be able to evaluate critically what they see, hear, and read. Also, with the huge amount of printed material available in all areas in this age of “information explosion” it is easy to feel overwhelmed. But often the information piled up on people’s desks and in their minds is of no use due to the enormous amount of it.

  6. Critical Mass

    CERN Multimedia

    AUTHOR|(CDS)2070299

    2017-01-01

    Critical Mass is a cycling event typically held on the last Friday of every month; its purpose is not usually formalized beyond the direct action of meeting at a set location and time and traveling as a group through city or town streets on bikes. The event originated in 1992 in San Francisco; by the end of 2003, the event was being held in over 300 cities around the world. At CERN it is held once a year in conjunction with the national Swiss campaing "Bike to work".

  7. Analyses of wrky18 wrky40 plants reveal critical roles of SA/EDS1 signaling and indole-glucosinolate biosynthesis for Golovinomyces orontii resistance and a loss-of resistance towards Pseudomonas syringae pv. tomato AvrRPS4.

    Science.gov (United States)

    Schön, Moritz; Töller, Armin; Diezel, Celia; Roth, Charlotte; Westphal, Lore; Wiermer, Marcel; Somssich, Imre E

    2013-07-01

    Simultaneous mutation of two WRKY-type transcription factors, WRKY18 and WRKY40, renders otherwise susceptible wild-type Arabidopsis plants resistant towards the biotrophic powdery mildew fungus Golovinomyces orontii. Resistance in wrky18 wrky40 double mutant plants is accompanied by massive transcriptional reprogramming, imbalance in salicylic acid (SA) and jasmonic acid (JA) signaling, altered ENHANCED DISEASE SUSCEPTIBILITY1 (EDS1) expression, and accumulation of the phytoalexin camalexin. Genetic analyses identified SA biosynthesis and EDS1 signaling as well as biosynthesis of the indole-glucosinolate 4MI3G as essential components required for loss-of-WRKY18 WRKY40-mediated resistance towards G. orontii. The analysis of wrky18 wrky40 pad3 mutant plants impaired in camalexin biosynthesis revealed an uncoupling of pre- from postinvasive resistance against G. orontii. Comprehensive infection studies demonstrated the specificity of wrky18 wrky40-mediated G. orontii resistance. Interestingly, WRKY18 and WRKY40 act as positive regulators in effector-triggered immunity, as the wrky18 wrky40 double mutant was found to be strongly susceptible towards the bacterial pathogen Pseudomonas syringae DC3000 expressing the effector AvrRPS4 but not against other tested Pseudomonas strains. We hypothesize that G. orontii depends on the function of WRKY18 and WRKY40 to successfully infect Arabidopsis wild-type plants while, in the interaction with P. syringae AvrRPS4, they are required to mediate effector-triggered immunity.

  8. Depletion of human micro-RNA miR-125b reveals that it is critical for the proliferation of differentiated cells but not for the down-regulation of putative targets during differentiation.

    Science.gov (United States)

    Lee, Yong Sun; Kim, Hak Kyun; Chung, Sangmi; Kim, Kwang-Soo; Dutta, Anindya

    2005-04-29

    Micro-RNAs are small non-coding RNAs that regulate target gene expression post-transcriptionally through base pairing with the target messenger RNA. Functional characterization of micro-RNAs awaits robust experimental methods to knock-down a micro-RNA as well as to assay its function in vivo. In addition to the recently developed method to sequester micro-RNA with 2'-O-methyl antisense oligonucleotide, we report that small interfering RNA against the loop region of a micro-RNA precursor can be used to deplete the micro-RNA. The depletion of miR-125b by this method had a profound effect on the proliferation of adult differentiated cancer cells, and this proliferation defect was rescued by co-transfected mature micro-RNA. This technique has unique advantages over the 2'-O-methyl antisense oligonucleotide and can be used to determine micro-RNA function, assay micro-RNAs in vivo, and identify the contribution of a predicted micro-RNA precursor to the pool of mature micro-RNA in a given cell. miR-125b and let-7 micro-RNAs are induced, whereas their putative targets, lin-28 and lin-41, are decreased during in vitro differentiation of Tera-2 or embryonic stem cells. Experimental increase or decrease of micro-RNA concentrations did not, however, affect the levels of the targets, a finding that is explained by the fact that the down-regulation of the targets appears to be mostly at the transcriptional level in these in vitro differentiation systems. Collectively these results reveal the importance of micro-RNA depletion strategies for directly determining micro-RNA function in vivo.

  9. The phase 2b HVTN 503/Phambili study test-of-concept HIV vaccine study, investigating a recombinant adenovirus type 5 HIV gag/pol/nef vaccine in South Africa: unblinded, long-term follow-up

    Science.gov (United States)

    Gray, Glenda E; Moodie, Zoe; Metch, Barbara; Gilbert, Peter B.; Bekker, Linda-Gail; Churchyard, Gavin; Nchabeleng, Maphoshane; Mlisana, Koleka; Laher, Fatima; Roux, Surita; Mngadi, Kathryn; Innes, Craig; Mathebula, Matsontso; Allen, Mary; McElrath, M Julie; Robertson, Michael; Kublin, James; Corey, Lawrence

    2014-01-01

    Background The Phambili study, conducted in South Africa amongst a predominantly heterosexual population, evaluated the efficacy of the MRK Ad5 gag/pol/nef subtype B HIV-1 preventive vaccine. Enrollment and vaccinations were stopped, participants unblinded, and follow-up extended when the Step study evaluating the same vaccine in the Americas, Caribbean and Australia was unblinded for non-efficacy with more HIV infections amongst vaccinee than placebo recipients [ZM1]. Extensive analyses over the complete follow-up period, most of which was unblinded, are reported. Methods Phambili participants were HIV-1 uninfected, sexually active men and women aged 18–35 years, followed for 3.5 years. HIV testing and risk reduction counseling occurred at weeks 0, 12, 30 and were switched to a 3 monthly schedule after unblinding. Cox proportional hazards models were used to estimate HIV-1 infection hazard ratios (HR) comparing vaccine to placebo recipients, overall and within subgroups. Long-term vaccine efficacy was evaluated in participants who were unblinded early in follow-up. Results Of the 801 participants enrolled (400 vaccine, 401 placebo), 112 (28%) received 1 vaccination, 259 (65%) 2 vaccinations and 29(7%) 3 vaccinations. More infections occurred in vaccinees (n=63) as compared to placebo (n=37) (adjusted HR (vaccine:placebo) 1.70, 95% CI 1.13–2.55, p = 0.01). We found no increase in infections with the number of vaccinations received and that the HRs did not differ by gender, circumcision, or Ad5 serostatus. Differences in risk behavior at baseline or during the study, or differential drop-out (p=0.40) are unlikely explanations for the increased rate of HIV-1 infections seen in vaccinees. Conclusion The increased HR of HIV-1 acquisition, irrespective of number of doses received, warrants further investigation to understand the biological mechanism. Further use of the Ad5 vector for HIV vaccines is not warranted PMID:24560541

  10. A Ricoeurian Approach to Rhetorical Criticism.

    Science.gov (United States)

    Warnick, Barbara

    1987-01-01

    Proposes an approach to rhetorical criticism that features the relationship between a message and a culturally distanced interpreter or critic. Applies Paul Ricoeur's critical approach to an interpretation of the Gettysburg Address to illustrate how hermeneutics reveals the underlying meaning and cultural significance of enduring texts. (NKA)

  11. Music critic Gustav Michel

    Directory of Open Access Journals (Sweden)

    Vasić Aleksandar N.

    2004-01-01

    oppose the National Theatre administrations when they decided to add operettas to its repertoire. Here he also differs from expert critics, for example Miloje Milojević or Petar Krstić - who led a real crusade against operetta. Michel paid scrupulous attention to correct diction, as an important part of the vocal technique. As a critic, Gustav Michel was inclined to relatively modern music. He was not strict in his judgments of Serbian performers’ and composers’ achievements; he always took account of very difficult conditions under which the Serbian people, after many centuries of the Turkish occupation, started its cultural and musical emancipation in the 19th century. (He was especially considerate towards novice musicians However his critical assessment of the genre status of the overture to the first Serbian opera, "Na uranku" ("At Dawn" by Stanislav Binički, revealed an incisive critic. The weak side of his critic lies in too general language not exact enough for characteristics of musical interpretations. However Gustav Michel was a witty and ironic writer, and his few articles marked the beginning of an expert and modern music critic in Serbia.

  12. Interpersonal criticism and the clergy.

    Science.gov (United States)

    Garner, Randy

    2013-03-01

    Though well-crafted criticism can provide valuable information for the recipient, the abundance of literature finds that criticism is overwhelmingly viewed negatively. The consequences of adverse interpersonal criticism were assessed by a focus group of randomly selected clergy members from a large ministerial alliance. Collectively the focus group revealed that interpersonal criticism can have deleterious vocational, psychological, and health consequence for those in the ministry. Clergy reported that criticism from parishioners, the public, denominational officials, and others adversely affect interpersonal relationships and can lead to stress, burnout, and early departure from the ministry. The participants indicated that current training and resources were inadequate to deal with the issue and further denominational support and seminary education was necessary.

  13. Critical theory and holocaust

    Directory of Open Access Journals (Sweden)

    Krstić Predrag

    2006-01-01

    Full Text Available In this paper the author is attempting to establish the relationship - or the lack of it - of the Critical Theory to the "Jewish question" and justification of perceiving signs of Jewish religious heritage in the thought of the representatives of this movement. The holocaust marked out by the name of "Auschwitz", is here tested as a point where the nature of this relationship has been decided. In this encounter with the cardinal challenge for the contemporary social theory, the particularity of the Frankfurt School reaction is here revealed through Adorno installing Auschwitz as unexpected but lawful emblem of the ending of the course that modern history has assumed. The critique of this "fascination" with Auschwitz, as well as certain theoretical pacification and measured positioning of the holocaust into discontinued plane of "unfinished" and continuation and closure of the valued project, are given through communicative-theoretical pre-orientation of Jürgen Habermas’s Critical Theory and of his followers. Finally, through the work of Detlev Claussen, it is suggested that in the youngest generation of Adorno’s students there are signs of revision to once already revised Critical Theory and a kind of defractured and differentiated return to the initial understanding of the decisiveness of the holocaust experience. This shift in the attitude of the Critical Theory thinkers to the provocation of holocaust is not, however, particularly reflected towards the status of Jews and their tradition, but more to the age old questioning and explanatory patterns for which they served as a "model". The question of validity of the enlightenment project, the nature of occidental rationalism, (nonexistence of historical theology and understanding of the identity and emancipation - describe the circle of problems around which the disagreement is concentrated in the social critical theory.

  14. Critical Pedagogy for Critical Mathematics Education

    Science.gov (United States)

    Tutak, Fatma Aslan; Bondy, Elizabeth; Adams, Thomasenia L.

    2011-01-01

    This article provides a brief introduction to critical pedagogy and further discussion on critical mathematics education. Critical mathematics education enables students to read the world with mathematics. Three emerging domains of mathematics education related to critical mathematics education are discussed in this manuscript: ethnomathematics,…

  15. Critical Pedagogy for Critical Mathematics Education

    Science.gov (United States)

    Tutak, Fatma Aslan; Bondy, Elizabeth; Adams, Thomasenia L.

    2011-01-01

    This article provides a brief introduction to critical pedagogy and further discussion on critical mathematics education. Critical mathematics education enables students to read the world with mathematics. Three emerging domains of mathematics education related to critical mathematics education are discussed in this manuscript: ethnomathematics,…

  16. Putting the Critical Back in Critical Infrastructure

    Science.gov (United States)

    2015-12-01

    CRITICAL BACK IN CRITICAL INFRASTRUCTURE by Bradford C. Mason December 2015 Thesis Advisor: Rudolph P. Darken Second Reader: Thomas Mackin...COVERED Master’s thesis 4. TITLE AND SUBTITLE PUTTING THE CRITICAL BACK IN CRITICAL INFRASTRUCTURE 5. FUNDING NUMBERS 6. AUTHOR(S) Bradford C. Mason...12b. DISTRIBUTION CODE A 13. ABSTRACT (maximum 200 words) In the context of national critical infrastructure security and resilience doctrine

  17. Cross-cultural perspectives on critical thinking.

    Science.gov (United States)

    Jenkins, Sheryl Daun

    2011-05-01

    The purpose of this cross-cultural study was to explore critical thinking among nurse scholars in Thailand and the United States. The study used qualitative methodology to examine how nurse scholars describe critical thinking in nursing. Nurse educators in Thailand and the United States were questioned concerning the following aspects of critical thinking: essential components; teaching and evaluation techniques; characteristics of critical thinkers; and the importance of a consensus definition for critical thinking in nursing. Their statements, which revealed both common and specific cultural aspects of critical thinking, were subjected to content analysis. Certain themes emerged that have not been widely discussed in the literature, including the link between staying calm and thinking critically, the assertion that happiness is an essential component of critical thinking, and the participants' nearly unanimous support for coming to a consensus definition of critical thinking for nursing.

  18. Introduction, Critical Text logy and Textual Criticism

    Directory of Open Access Journals (Sweden)

    فرزاد قائمی

    2013-06-01

    Full Text Available Asadi’s Shahnameh is a great epic consisting of twenty-four thousand distiches and is attributed to Asadi or another poet of the same nickname. This work was created in the same line of development as Ferdowsi’s Shahnameh. The main theme is the old campaign of Soleymān to Iran to confront with Rostam and Keykhosrow and to repeat the pattern of Rostam’s battles with his children in a state of anonymity. The text structure is episodic with numerous central characters. The narratives are for the most part derived from oral literature. Textual evidence demonstrates that the poet is Shiite. The narrative content, chronogram as well as the literary and linguistic style of one of the manuscripts reveal that the text was written in the ninth century (probably 809 A.H.. The article first introduces the text and the origin of its narratives in oral literature; it then proceeds with the study of the narrative structure of the epic using three available manuscripts dating back to the thirteenth and fourteenth centuries (A.H.. Textology and Textual Criticism have been employed as the research methodology. The literary and linguistic features of the text have also been examined at three levels: lexical, syntactic and rhetorical.

  19. Quantum Criticality via Magnetic Branes

    Science.gov (United States)

    D'Hoker, Eric; Kraus, Per

    Holographic methods are used to investigate the low temperature limit, including quantum critical behavior, of strongly coupled 4-dimensional gauge theories in the presence of an external magnetic field, and finite charge density. In addition to the metric, the dual gravity theory contains a Maxwell field with Chern-Simons coupling. In the absence of charge, the magnetic field induces an RG flow to an infrared {AdS}3 × {R}2 geometry, which is dual to a 2-dimensional CFT representing strongly interacting fermions in the lowest Landau level. Two asymptotic Virasoro algebras and one chiral Kac-Moody algebra arise as emergent symmetries in the IR. Including a nonzero charge density reveals a quantum critical point when the magnetic field reaches a critical value whose scale is set by the charge density. The critical theory is probed by the study of long-distance correlation functions of the boundary stress tensor and current. All quantities of major physical interest in this system, such as critical exponents and scaling functions, can be computed analytically. We also study an asymptotically AdS 6 system whose magnetic field induced quantum critical point is governed by an IR Lifshitz geometry, holographically dual to a D=2+1 field theory. The behavior of these holographic theories shares important similarities with that of real world quantum critical systems obtained by tuning a magnetic field, and may be relevant to materials such as Strontium Ruthenates.

  20. Media Literacy Function in Critical Blogs

    Science.gov (United States)

    Fedorov, Alexander; Levitskaya, Anastasia

    2015-01-01

    The Internet is widely recognized as playing an important role in facilitating education on a range of issues, including media literacy. Analyzing the media critical activity of contemporary Russian bloggers, the authors of the article reveal the following reasons for popularity or, on the contrary, unpopularity of blogger's media criticism:…

  1. Critical Thinking in the Business Classroom

    Science.gov (United States)

    Reid, Joanne R.; Anderson, Phyllis R.

    2012-01-01

    A minicourse in critical thinking was implemented to improve student outcomes in two sessions of a senior-level business course at a Midwestern university. Statistical analyses of two quantitative assessments revealed significant improvements in critical thinking skills. Improvements in student outcomes in case studies and computerized business…

  2. Critical Thinking in the Business Classroom

    Science.gov (United States)

    Reid, Joanne R.; Anderson, Phyllis R.

    2012-01-01

    A minicourse in critical thinking was implemented to improve student outcomes in two sessions of a senior-level business course at a Midwestern university. Statistical analyses of two quantitative assessments revealed significant improvements in critical thinking skills. Improvements in student outcomes in case studies and computerized business…

  3. Nuclear criticality safety guide

    Energy Technology Data Exchange (ETDEWEB)

    Pruvost, N.L.; Paxton, H.C. [eds.

    1996-09-01

    This technical reference document cites information related to nuclear criticality safety principles, experience, and practice. The document also provides general guidance for criticality safety personnel and regulators.

  4. A kinematic study of critical and non-critical articulators in emotional speech production.

    Science.gov (United States)

    Kim, Jangwon; Toutios, Asterios; Lee, Sungbok; Narayanan, Shrikanth S

    2015-03-01

    This study explores one aspect of the articulatory mechanism that underlies emotional speech production, namely, the behavior of linguistically critical and non-critical articulators in the encoding of emotional information. The hypothesis is that the possible larger kinematic variability in the behavior of non-critical articulators enables revealing underlying emotional expression goal more explicitly than that of the critical articulators; the critical articulators are strictly controlled in service of achieving linguistic goals and exhibit smaller kinematic variability. This hypothesis is examined by kinematic analysis of the movements of critical and non-critical speech articulators gathered using eletromagnetic articulography during spoken expressions of five categorical emotions. Analysis results at the level of consonant-vowel-consonant segments reveal that critical articulators for the consonants show more (less) peripheral articulations during production of the consonant-vowel-consonant syllables for high (low) arousal emotions, while non-critical articulators show less sensitive emotional variation of articulatory position to the linguistic gestures. Analysis results at the individual phonetic targets show that overall, between- and within-emotion variability in articulatory positions is larger for non-critical cases than for critical cases. Finally, the results of simulation experiments suggest that the postural variation of non-critical articulators depending on emotion is significantly associated with the controls of critical articulators.

  5. A Critical Humanist Curriculum

    Science.gov (United States)

    Magill, Kevin; Rodriguez, Arturo

    2015-01-01

    This essay is a critical humanist discussion of curriculum; a departure from the technicist view of education [education meant to support a global capitalist economy] and an analysis of curriculum considering critical humanism, political economy and critical race theory among other modes of critical analysis and inquiry. Our discussion supports a…

  6. Criticality Analysis of GTPPS

    Directory of Open Access Journals (Sweden)

    Asis Sarkar

    2011-10-01

    Full Text Available The paper is concerned with the study of criticality analysis of components of Gas Turbine Power Plant Systems (GTPPS and the failures occurring in the plant. Failure mode and effect and criticality analysis (FMECA is carried out to estimate the criticality number for different components and failure modes. In addition the failure effects, higher effects and end effectsare incorporated in the final FMECA sheet. The criticality resultscompensating provision will highlight possible ways to tackle thefailures economically. The findings in this Paper are (1 criticality index of the components (2 Critical failures (3 compensating provision of critical failure.

  7. A Critical Look into Critical Pedagogy

    Science.gov (United States)

    Pishghadam, Reza; Meidani, Elham Naji

    2012-01-01

    In line with postmodern philosophy, critical pedagogy has gained considerable importance and has become a valuable educational goal. The purpose of this study is to dig into the effects of critical pedagogy in a modernist educational system. To this aim, 15 Iranian university students were asked to write down their feelings at the end of a course…

  8. A Critical Look into Critical Pedagogy

    Science.gov (United States)

    Pishghadam, Reza; Meidani, Elham Naji

    2012-01-01

    In line with postmodern philosophy, critical pedagogy has gained considerable importance and has become a valuable educational goal. The purpose of this study is to dig into the effects of critical pedagogy in a modernist educational system. To this aim, 15 Iranian university students were asked to write down their feelings at the end of a course…

  9. Critical Approaches to Critical Pedagogy in Greece

    Science.gov (United States)

    Grollios, George; Liambas, Anastassios

    2015-01-01

    This paper is an initial review of the presentations and uses of critical pedagogy in Greek educational literature since the mid-1980s. These have appeared in the form of three books and an edited volume (translated in Greek), all written by American critical educators, as well as in texts produced by Greek educators who have either written the…

  10. From Critical Theory to Critical Hermeneutics

    National Research Council Canada - National Science Library

    Øjvind Larsen

    2014-01-01

    From their beginning in the 1930s, critical theory and the Frankfurt school had their focus on a critique of disturbed social relations in western society dominated by totalitarian political regimes...

  11. From Critical Theory to Critical Hermeneutics

    OpenAIRE

    Øjvind Larsen

    2014-01-01

    From their beginning in the 1930s, critical theory and the Frankfurt school had their focus on a critique of disturbed social relations in western society dominated by totalitarian political regimes like Stalinism, Fascism, Nazism, and by capitalism as an oppressive and destructive economic system and culture. Now, 80 years later, this has all become history and thus it is time to leave the concept of critical theory behind us, and instead bring the concept of critique to a broader theoretica...

  12. Surgical Critical Care Initiative

    Data.gov (United States)

    Federal Laboratory Consortium — The Surgical Critical Care Initiative (SC2i) is a USU research program established in October 2013 to develop, translate, and validate biology-driven critical care....

  13. Interface localization near criticality

    CERN Document Server

    Delfino, Gesualdo

    2016-01-01

    The theory of interface localization in near-critical planar systems at phase coexistence is formulated from first principles. We show that mutual delocalization of two interfaces, amounting to interfacial wetting, occurs when the bulk correlation length critical exponent $\

  14. California Condor Critical Habitat

    Data.gov (United States)

    California Department of Resources — These Data identify (in general) the areas where critical habitat for the California Condor occur. Critical habitat for the species consists of the following 10...

  15. Critical dynamics near QCD critical point

    CERN Document Server

    Minami, Yuki

    2012-01-01

    In this thesis, we study the critical dynamics near the QCD critical point. Near the critical point, the relevant modes for the critical dynamics are identified as the hydrodynamic modes. Thus, we first study the linear dynamics of them by the relativistic hydrodynamics. We show that the thermal diffusion mode is the most relevant mode, whereas the sound mode is suppressed around the critical point. We also find that the Landau equation, which is believed to be an acausal hydrodynamic equation, has no problem to describe slowly varying fluctuations. Moreover, we find that the Israel-Stewart equation, which is a causal one, gives the same result as the Landau equation gives in the long-wavelength region. Next, we study the nonlinear dynamics of the hydrodynamic modes by the nonlinear Langevin equation and the dynamic renormalization group (RG). In the vicinity of the critical point, the usual hydrodynamics breaks down by large fluctuations. Thus, we must consider the nonlinear Langevin equation. We construct t...

  16. Toward critical bioethics.

    Science.gov (United States)

    Árnason, Vilhjálmur

    2015-04-01

    This article deals with the question as to what makes bioethics a critical discipline. It considers different senses of criticism and evaluates their strengths and weaknesses. A primary method in bioethics as a philosophical discipline is critical thinking, which implies critical evaluation of concepts, positions, and arguments. It is argued that the type of analytical criticism that restricts its critical role to critical thinking of this type often suffers from other intellectual flaws. Three examples are taken to demonstrate this: premature criticism, uncritical self-understanding of theoretical assumptions, and narrow framing of bioethical issues. Such flaws can lead both to unfair treatment of authors and to uncritical discussion of topics. In this context, the article makes use of Häyry's analysis of different rationalities in bioethical approaches and argues for the need to recognize the importance of communicative rationality for critical bioethics. A radically different critical approach in bioethics, rooted in social theory, focuses on analyses of power relations neglected in mainstream critical thinking. It is argued that, although this kind of criticism provides an important alternative in bioethics, it suffers from other shortcomings that are rooted in a lack of normative dimensions. In order to complement these approaches and counter their shortcomings, there is a need for a bioethics enlightened by critical hermeneutics. Such hermeneutic bioethics is aware of its own assumptions, places the issues in a wide context, and reflects critically on the power relations that stand in the way of understanding them. Moreover, such an approach is dialogical, which provides both a critical exercise of speech and a normative dimension implied in the free exchange of reasons and arguments. This discussion is framed by Hedgecoe's argument that critical bioethics needs four elements: to be empirically rooted, theory challenging, reflexive, and politely skeptical.

  17. Reimagining Critical Theory

    Science.gov (United States)

    Rexhepi, Jevdet; Torres, Carlos Alberto

    2011-01-01

    This paper discusses Critical Theory, a model of theorizing in the field of the political sociology of education. We argue for a "reimagined" Critical Theory to herald an empowering, liberatory education that fosters curiosity and critical thinking, and a means for successful bottom-up, top-down political engagement. We present arguments at a…

  18. Reimagining Critical Theory

    Science.gov (United States)

    Rexhepi, Jevdet; Torres, Carlos Alberto

    2011-01-01

    This paper discusses Critical Theory, a model of theorizing in the field of the political sociology of education. We argue for a "reimagined" Critical Theory to herald an empowering, liberatory education that fosters curiosity and critical thinking, and a means for successful bottom-up, top-down political engagement. We present arguments…

  19. Foundations for Critical Thinking

    Science.gov (United States)

    Bers, Trudy; Chun, Marc; Daly, William T.; Harrington, Christine; Tobolowsky, Barbara F.

    2015-01-01

    "Foundations for Critical Thinking" explores the landscape of critical-thinking skill development and pedagogy through foundational chapters and institutional case studies involving a range of students in diverse settings. By establishing a link between active learning and improved critical thinking, this resource encourages all higher…

  20. Reimagining Critical Theory

    Science.gov (United States)

    Rexhepi, Jevdet; Torres, Carlos Alberto

    2011-01-01

    This paper discusses Critical Theory, a model of theorizing in the field of the political sociology of education. We argue for a "reimagined" Critical Theory to herald an empowering, liberatory education that fosters curiosity and critical thinking, and a means for successful bottom-up, top-down political engagement. We present arguments…

  1. International critical perspectives

    NARCIS (Netherlands)

    Sambrook, S.A.; Poell, R.F.

    2014-01-01

    The Problem Critical perspectives on human resource development (HRD) have emerged, across Europe and America, hailed as the future of the field. However, we note the paucity of critical perspectives globally, the problematic dominance of critical HRD activities in Western sites of theory and

  2. Criticism, Visible and Invisible.

    Science.gov (United States)

    Frye, Northrop

    1964-01-01

    The central activity of literary criticism, the understanding of literature, is related to the process of establishing a context for the works of literature being studied. Choosing not to discuss the factual elements of literary criticism, the author clarifies and concentrates on the "lower" and "upper" limits of criticism. While the "lower" limit…

  3. Creating a Critical Thinker

    Science.gov (United States)

    Piergiovanni, Polly R.

    2014-01-01

    A college education is expected to improve students' critical thinking skills. Keeping students active in class--through writing activities and class discussion--has been shown to help students think critically. In this article, creative hands-on activities, which are common in engineering courses, are shown to improve students' critical thinking…

  4. Reconceptualising Critical Digital Literacy

    Science.gov (United States)

    Pangrazio, Luciana

    2016-01-01

    While it has proved a useful concept during the past 20 years, the notion of "critical digital literacy" requires rethinking in light of the fast-changing nature of young people's digital practices. This paper contrasts long-established notions of "critical digital literacy" (based primarily around the critical consumption of…

  5. Reconceptualising Critical Digital Literacy

    Science.gov (United States)

    Pangrazio, Luciana

    2016-01-01

    While it has proved a useful concept during the past 20 years, the notion of "critical digital literacy" requires rethinking in light of the fast-changing nature of young people's digital practices. This paper contrasts long-established notions of "critical digital literacy" (based primarily around the critical consumption of…

  6. Creating a Critical Thinker

    Science.gov (United States)

    Piergiovanni, Polly R.

    2014-01-01

    A college education is expected to improve students' critical thinking skills. Keeping students active in class--through writing activities and class discussion--has been shown to help students think critically. In this article, creative hands-on activities, which are common in engineering courses, are shown to improve students' critical thinking…

  7. A Critical Analysis of the Conceptualisation of "Coaching Philosophy"

    Science.gov (United States)

    Cushion, Christopher; Partington, Mark

    2016-01-01

    The aim of this paper was to critically review existing literature relating to, and critically analyse current conceptualisations of, "coaching philosophy." The review reveals a bewildering approach to definitions, terms and frameworks that have limited explanation and reveal a lack of conceptual clarity. It is argued that rather than…

  8. A Critical Analysis of the Conceptualisation of "Coaching Philosophy"

    Science.gov (United States)

    Cushion, Christopher; Partington, Mark

    2016-01-01

    The aim of this paper was to critically review existing literature relating to, and critically analyse current conceptualisations of, "coaching philosophy." The review reveals a bewildering approach to definitions, terms and frameworks that have limited explanation and reveal a lack of conceptual clarity. It is argued that rather than…

  9. Critical Reflectivity and the Development of New Culturally Relevant Teachers

    Science.gov (United States)

    Durden, Tonia R.; Truscott, Diane M.

    2013-01-01

    Three case studies present how preservice teachers use reflections while learning to teach. Interviews and document analysis reveal that critical reflections evidence greater understanding of culturally relevant pedagogy and offer a platform for critical consciousness. Using critical reflectivity to develop teachers' understandings of culturally…

  10. From Critical Theory to Critical Hermeneutics

    Directory of Open Access Journals (Sweden)

    Øjvind Larsen

    2014-06-01

    Full Text Available From their beginning in the 1930s, critical theory and the Frankfurt school had their focus on a critique of disturbed social relations in western society dominated by totalitarian political regimes like Stalinism, Fascism, Nazism, and by capitalism as an oppressive and destructive economic system and culture. Now, 80 years later, this has all become history and thus it is time to leave the concept of critical theory behind us, and instead bring the concept of critique to a broader theoretical framework like hermeneutics. This allows the possibility of retaining the theoretical intentions of the old Frankfurt school and at the same time there will be no boundaries by specific dominant theoretical perspectives. In this paper, such a framework for a critical hermeneutics is discussed on the basis of Weber’s, Gadamer’s, and Habermas’ theories on hermeneutics within the social sciences.

  11. Media Literacy Function in Critical Blogs

    Directory of Open Access Journals (Sweden)

    Alexander Fedorov

    2015-04-01

    Full Text Available The Internet is widely recognized as playing an important role in facilitating education on a range of issues, including media literacy. Analyzing the media critical activity of contemporary Russian bloggers, the authors of the article reveal the following reasons for popularity or, on the contrary, unpopularity of blogger's media criticism: targeted orientation, emotional charge, entertainment nature, duration, interactiveness, multimedia mode, simplicity/complexity of the language of a media text, the level of conformity.

  12. Critical Psychologies for Critical Health Literacies

    Science.gov (United States)

    Corcoran, Tim

    2014-01-01

    Health education is largely informed by psychological theories and practices that pursue reductionist views of people learning. However, critical attention is moving to understand health in ways that reconsider relationships to context and the forms of life within which everyday living takes place. This shift is apparent in theoretical…

  13. Critical Psychologies for Critical Health Literacies

    Science.gov (United States)

    Corcoran, Tim

    2014-01-01

    Health education is largely informed by psychological theories and practices that pursue reductionist views of people learning. However, critical attention is moving to understand health in ways that reconsider relationships to context and the forms of life within which everyday living takes place. This shift is apparent in theoretical…

  14. Critical incident reporting systems.

    Science.gov (United States)

    Ahluwalia, Jag; Marriott, Lin

    2005-02-01

    Approximately 10% of all hospital admissions are complicated by critical incidents in which harm is caused to the patient - this amounts to more than 850,000 incidents annually. Critical incident reporting (CIR) systems refer to the structured reporting, collation and analysis of such incidents. This article describes the attributes required for an effective CIR system. Example neonatal trigger events and a management pathway for handling a critical incident report are described. The benefits and limitations of CIR systems, reactive and prospective approaches to the analysis of actual or potential critical incidents and the assessment of risk are also reviewed. Individual human error is but one contributor in the majority of critical incidents. Recognition of this and the fostering of an organisational culture that views critical incident reports as an opportunity to learn and to improve future patient care is vital if CIR systems are to be effective.

  15. Vulnerability of critical infrastructures : identifying critical nodes.

    Energy Technology Data Exchange (ETDEWEB)

    Cox, Roger Gary; Robinson, David Gerald

    2004-06-01

    The objective of this research was the development of tools and techniques for the identification of critical nodes within critical infrastructures. These are nodes that, if disrupted through natural events or terrorist action, would cause the most widespread, immediate damage. This research focuses on one particular element of the national infrastructure: the bulk power system. Through the identification of critical elements and the quantification of the consequences of their failure, site-specific vulnerability analyses can be focused at those locations where additional security measures could be effectively implemented. In particular, with appropriate sizing and placement within the grid, distributed generation in the form of regional power parks may reduce or even prevent the impact of widespread network power outages. Even without additional security measures, increased awareness of sensitive power grid locations can provide a basis for more effective national, state and local emergency planning. A number of methods for identifying critical nodes were investigated: small-world (or network theory), polyhedral dynamics, and an artificial intelligence-based search method - particle swarm optimization. PSO was found to be the only viable approach and was applied to a variety of industry accepted test networks to validate the ability of the approach to identify sets of critical nodes. The approach was coded in a software package called Buzzard and integrated with a traditional power flow code. A number of industry accepted test networks were employed to validate the approach. The techniques (and software) are not unique to power grid network, but could be applied to a variety of complex, interacting infrastructures.

  16. Surveillance and Critical Theory

    Directory of Open Access Journals (Sweden)

    Christian Fuchs

    2015-09-01

    Full Text Available In this comment, the author reflects on surveillance from a critical theory approach, his involvement in surveillance research and projects, and the status of the study of surveillance. The comment ascertains a lack of critical thinking about surveillance, questions the existence of something called “surveillance studies” as opposed to a critical theory of society, and reflects on issues such as Edward Snowden’s revelations, and Foucault and Marx in the context of surveillance.

  17. Extending Critical Performativity

    DEFF Research Database (Denmark)

    Spicer, André; Alvesson, Mats; Kärreman, Dan

    2016-01-01

    In this article we extend the debate about critical performativity. We begin by outlining the basic tenets of critical performativity and how this has been applied in the study of management and organization. We then address recent critiques of critical performance. We note these arguments suffer...... from an undue focus on intra-academic debates; engage in author-itarian theoretical policing; feign relevance through symbolic radicalism; and repackage common sense. We take these critiques as an opportunity to offer an extended model of critical performativity that involves focusing on issues...

  18. Constructing criticality by classification

    DEFF Research Database (Denmark)

    Machacek, Erika

    2017-01-01

    This paper explores the role of expertise, the nature of criticality, and their relationship to securitisation as mineral raw materials are classified. It works with the construction of risk along the liberal logic of security to explore how "key materials" are turned into "critical materials......, legitimizing a criticality discourse.Specifically, the paper introduces a typology delineating the inferences made by the experts from their produced recommendations in the classification of rare earth element criticality. The paper argues that the classification is a specific process of constructing risk...

  19. Furthering critical institutionalism

    NARCIS (Netherlands)

    Cleaver, Frances; Koning, De Jessica

    2015-01-01

    This special issue furthers the study of natural resource management from a critical institutional perspective. Critical institutionalism (CI) is a contemporary body of thought that explores how institutions dynamically mediate relationships between people, natural resources and society. It focus

  20. Critical Care Team

    Science.gov (United States)

    ... Patients and Families > About Critical Care > Team Tweet Team Page Content ​The critical care team is a group of specially trained caregivers who ... help very ill patients get better. The care team often teach the patient and family strategies that ...

  1. Creating a Critical Thinker

    Science.gov (United States)

    Piergiovanni, Polly R.

    2014-01-01

    A college education is expected to improve students' critical thinking skills. Keeping students active in class--through writing activities and class discussion--has been shown to help students think critically. In this article, creative hands-on activities, which are common in engineering courses, are shown to improve students' critical…

  2. Against Critical Thinking Pedagogy

    Science.gov (United States)

    Hayes, David

    2015-01-01

    Critical thinking pedagogy is misguided. Ostensibly a cure for narrowness of thought, by using the emotions appropriate to conflict, it names only one mode of relation to material among many others. Ostensibly a cure for fallacies, critical thinking tends to dishonesty in practice because it habitually leaps to premature ideas of what the object…

  3. Critical Literacy: Foundational Notes

    Science.gov (United States)

    Luke, Allan

    2012-01-01

    The term "critical literacy" refers to use of the technologies of print and other media of communication to analyze, critique, and transform the norms, rule systems, and practices governing the social fields of everyday life (A. Luke, 2004). Since Freire's (1970) educational projects in Brazil, approaches to critical literacy have been developed…

  4. Critical Utopian Action Research

    DEFF Research Database (Denmark)

    Nielsen, Birger Steen; Nielsen, Kurt Aagaard

    2016-01-01

    The specific concept of critical utopian action research is presented and discussed, as to its origin, use and potentials. The inspiration from Robert Jungk and his future creating workshops is elaborated.......The specific concept of critical utopian action research is presented and discussed, as to its origin, use and potentials. The inspiration from Robert Jungk and his future creating workshops is elaborated....

  5. Critical Literacy: Foundational Notes

    Science.gov (United States)

    Luke, Allan

    2012-01-01

    The term "critical literacy" refers to use of the technologies of print and other media of communication to analyze, critique, and transform the norms, rule systems, and practices governing the social fields of everyday life (A. Luke, 2004). Since Freire's (1970) educational projects in Brazil, approaches to critical literacy have been…

  6. Rethinking Critical Thinking

    Science.gov (United States)

    Downs, Christopher J.

    2008-01-01

    Critical thinking is of primary importance in higher education, yet the concept remains slippery and the skill elusive. The author argues that most current critical thinking textbooks are out of line with the seminal work of John Dewey. Rather than logical argument and justification, it is suggested that carefulness, open-mindedness and creativity…

  7. Critical Teaching and Learning.

    Science.gov (United States)

    Young, R. E.

    1988-01-01

    This essay begins with a description of Jurgen Habermas's theory of communicative action, emphasizing aspects which relate to a critical theory of teaching/learning. Existing theory and research is reviewed. A critical theory of pedagogy as a form of communicative action or interaction is presented. (MT)

  8. Critical Discourse Analysis

    Institute of Scientific and Technical Information of China (English)

    杜梅香

    2006-01-01

    This paper is about the discourse analysis and illustrate the approach to analysis the Critical discourses and the discourses about the educational situation of China. And it also includes the condensed theoretical support of the Critical discourse analysis and analysis of the sample I of the discourses between an illiterate person and the literate.

  9. Critical Literacy: Foundational Notes

    Science.gov (United States)

    Luke, Allan

    2012-01-01

    The term "critical literacy" refers to use of the technologies of print and other media of communication to analyze, critique, and transform the norms, rule systems, and practices governing the social fields of everyday life (A. Luke, 2004). Since Freire's (1970) educational projects in Brazil, approaches to critical literacy have been…

  10. Critical Schwinger Pair Production.

    Science.gov (United States)

    Gies, Holger; Torgrimsson, Greger

    2016-03-04

    We investigate Schwinger pair production in spatially inhomogeneous electric backgrounds. A critical point for the onset of pair production can be approached by fields that marginally provide sufficient electrostatic energy for an off-shell long-range electron-positron fluctuation to become a real pair. Close to this critical point, we observe features of universality which are analogous to continuous phase transitions in critical phenomena with the pair-production rate serving as an order parameter: electric backgrounds can be subdivided into universality classes and the onset of pair production exhibits characteristic scaling laws. An appropriate design of the electric background field can interpolate between power-law scaling, essential Berezinskii-Kosterlitz-Thouless-type scaling, and a power-law scaling with log corrections. The corresponding critical exponents only depend on the large-scale features of the electric background, whereas the microscopic details of the background play the role of irrelevant perturbations not affecting criticality.

  11. Predictability of Critical Transitions

    CERN Document Server

    Zhang, Xiaozhu; Hallerberg, Sarah

    2015-01-01

    Critical transitions in multistable systems have been discussed as models for a variety of phenomena ranging from the extinctions of species to socio-economic changes and climate transitions between ice-ages and warm-ages. From bifurcation theory we can expect certain critical transitions to be preceded by a decreased recovery from external perturbations. The consequences of this critical slowing down have been observed as an increase in variance and autocorrelation prior to the transition. However especially in the presence of noise it is not clear, whether these changes in observation variables are statistically relevant such that they could be used as indicators for critical transitions. In this contribution we investigate the predictability of critical transitions in conceptual models. We study the the quadratic integrate-and-fire model and the van der Pol model, under the influence of external noise. We focus especially on the statistical analysis of the success of predictions and the overall predictabil...

  12. Critical illness myopathy.

    Science.gov (United States)

    Latronico, Nicola; Tomelleri, Giuliano; Filosto, Massimiliano

    2012-11-01

    To describe the incidence, major risk factors, and the clinical, electrophysiological, and histological features of critical illness myopathy (CIM). Major pathogenetic mechanisms and long-term consequences of CIM are also reviewed. CIM is frequently associated with critical illness polyneuropathy (CIP), and may have a relevant impact on patients' outcome. CIM has an earlier onset than CIP, and recovery is faster. Loss of myosin filaments on muscle biopsy is important to diagnose CIM, and has a good prognosis. Critical illness, use of steroids, and immobility concur in causing CIM. A rationale diagnostic approach to CIM using clinical, electrophysiological, and muscle biopsy investigations is important to plan adequate therapy and to predict recovery.

  13. Green Turtle Critical Habitat

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — These data represent the critical habitat for green turtle as designated by Federal Register Vol. 63, No. 46701, September 2, 1998, Rules and Regulations.

  14. Designated Critical Habitat

    Data.gov (United States)

    Kansas Data Access and Support Center — Critical habitats include those areas documented as currently supporting self-sustaining populations of any threatened or endangered species of wildlife as well as...

  15. Art Criticism and Reading.

    Science.gov (United States)

    Feldman, Edmund Burke; Woods, Don

    1981-01-01

    The authors review a body of theory and accumulating evidence which suggests that critical study of the arts facilitates the development of cognitive skills, including those essential to reading. (Author/SJL)

  16. Green Turtle Critical Habitat

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — These data represent the critical habitat for green turtle as designated by Federal Register Vol. 63, No. 46701, September 2, 1998, Rules and Regulations.

  17. Right Whale Critical Habitat

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These data represent the critical habitat for Right Whale as designated by Federal Register Vol. 59, No. 28805, May 19, 1993, Rules and Regulations.

  18. Critical Limb Ischemia (CLI)

    Science.gov (United States)

    ... Buerger’s Disease Carotid Artery Disease Chronic Venous Insufficiency Congenital Vascular Malformation Critical Limb Ischemia (CLI) Deep Vein Thrombosis (DVT) Diabetes and Vascular Disease Fibromuscular Dysplasia High Blood Pressure and Vascular Disease Kidney Failure ...

  19. Critical Schwinger pair production

    CERN Document Server

    Gies, Holger

    2015-01-01

    We investigate Schwinger pair production in spatially inhomogeneous electric backgrounds. A critical point for the onset of pair production can be approached by fields that marginally provide sufficient electrostatic energy for an off-shell long-range electron-positron fluctuation to become a real pair. Close to this critical point, we observe features of universality which are analogous to continuous phase transitions in critical phenomena with the pair-production rate serving as an order parameter: electric backgrounds can be subdivided into universality classes and the onset of pair production exhibits characteristic scaling laws. An appropriate design of the electric background field can interpolate between power-law scaling, essential BKT-type scaling and a power-law scaling with log corrections. The corresponding critical exponents only depend on the large-scale features of the electric background, whereas the microscopic details of the background play the role of irrelevant perturbations not affecting ...

  20. Wittgenstein and Critical Reasoning

    Directory of Open Access Journals (Sweden)

    Richard Shusterman

    2016-06-01

    Full Text Available In his paper Wittgenstein and Critical Reasoning, Richard Shusterman analyses the influence which Wittgensteinian aesthetics has had on contemporary criticism. Concentrating primarily on the second phase of Wittgenstein’s philosophy, Shusterman first demonstrates how the idea of language games and related concepts undermined both essentialist deductive and quasi-scientific inductive models of criticism. In place of these two approaches, Shusterman argues, Wittgensteinian aesthetics offers a new one, which he describes as perceptual-persuasive. In the next step, however, Shusterman observes that Wittgenstein’s followers often commit the very fallacy that Wittgenstein wanted to avoid, by imposing the perceptual persuasive model as the only valid solution for criticism. However, in order to be more in line with Wittgensteinian aesthetics, he argues, we should rather recognise the plurality of possibilities – perceptual-persuasive, deductive or inductive – whose choice depends on the language game currently played.

  1. Green Turtle Critical Habitat

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These data represent the critical habitat for green turtle as designated by Federal Register Vol. 63, No. 46701, September 2, 1998, Rules and Regulations.

  2. Critical Habitat Designations

    Data.gov (United States)

    Department of Homeland Security — The Endangered Species Act (ESA) requires the Federal government to designate 'critical habitat' for any species it lists under the ESA. This dataset combines both...

  3. Johnsons Seagrass Critical Habitat

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These data represent the critical habitat for Johnson's Seagrass as designated by Federal Register Vol. 65, No. 66, Wednesday, April 5, 2000, Rules and Regulations.

  4. Smalltooth Sawfish Critical Habitat

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These data represent the critical habitat for smalltooth sawfish (Pristis pectinatat) as designated by 74 FR 45353, September 2, 2009, Rules and Regulations.

  5. [Severe infection in critical emergency care].

    Science.gov (United States)

    Matsuda, Naoyuki; Takatani, Yudai; Higashi, Tomoko; Inaba, Masato; Ejima, Tadashi

    2016-02-01

    In the emergency and critical care medicine, infection is easy to merge to various basic conditions and diseases. In the social structure aging in critical care, the immune weakness was revealed as the result of severe infection and septic shock in the reduced function of neutrophils and lymphocytes. In the life-saving emergency care, cardiovascular diseases, diabetes, chronic renal failure and lever dysfunction are often observed, and the underlying diseases have the foundation of biological invasion after a first inflammatory attack of surgery, trauma, burn, and systemic injury. It will be placed into a susceptible situation such as artificial respiratory management. In this review, we discussed severe infection in emergency and critical care. It is necessary to pay attention to the drug resistance bacterias in own critical care setting by trends.

  6. Heat reveals faults

    Energy Technology Data Exchange (ETDEWEB)

    Weinreich, Bernhard [Solarschmiede GmbH, Muenchen (Germany). Engineering Dept.

    2010-07-01

    Gremlins cannot hide from the all-revealing view of a thermographic camera, whereby it makes no difference whether it is a roof-mounted system or a megawatt-sized farm. Just as diverse are the range of faults that, with the growing level of expertise, can now be detected and differentiated with even greater detail. (orig.)

  7. Danto and Art Criticism

    Directory of Open Access Journals (Sweden)

    Cynthia Freeland

    2008-01-01

    Full Text Available In this article I examine the relationship between Arthur Danto's philosophy of art and his practice of art criticism. Danto has said that he included many actual examples of discussions of art in The Transfiguration of the Commonplace because of the feeling that, previously, philosophers had theorized about art in a vacuum. And since the time of publishing that book, he has written on a wide variety of both historical and contemporary artists and art practices. Danto's philosophy of art commits him to an account of the practice of art criticism as interpretation. However, I question whether the Danto-esque interpretive essay can serve as an adequate model for art criticism. My primary claim is that art criticism must include a more strongly evaluative element than Danto's theory leaves room for, since on his view, the critic primarily explains meaning by examining how it is embodied in a work. This leaves open the question of which meanings count as valuable or important. In his more recent work Danto has explored a "Hegelian" view that art is primarily about art, but this view too does not allow for art to be evaluated or "criticized" on the basis of whether or how well it tackles the more profound questions of meaning.

  8. The Critical Bricolage

    Directory of Open Access Journals (Sweden)

    Kim McMillan

    2015-10-01

    Full Text Available Change in health care has become rapid and continuous. Much decision-making processes guiding change management are derived from organizational literature which is heavily reflective of managerial perspectives. These perspectives represent and aim to serve only a small subgroup of organizational members. However, change is complex, fragile, and has higher rates of success and sustainability when change management strategies reflect a multitude of organizational voices. There is a dearth of literature exploring the intersect between organizational and nursing discourses on the subject of rapid and continuous change in health care. Multitheoretical, multimethodological, and multidisciplinary informed approaches to methodological decision making are needed to link organizational and nursing discourses in ways that will offer alternative perspectives on the subject of change. Furthermore, critically guided multitheoretical, multimethodological, and multidisciplinary methodologies are timely and important in organizational research. Critically guided research seeks to analyze taken-for-granted assumptions and institutionalized practices seeking alternative perspectives and alternative sources of organizational knowledge. Exploring alternative perspectives from a critical lens recognizes the impact predominant discursive influences have on change management and the subsequent impact on organizational members’ working lives. This article will explore how Kincheloe’s discussions of the critical bricolage serve to support combining critical organizational methodologies (guided by Alvesson and Deetz with a voice-centered relational method of data analysis (guided by Brown and Gilligan to create a critical interpretive methodology that explores the voices of nurses as they experience rapid and continuous change in health care.

  9. Furthering critical institutionalism

    Directory of Open Access Journals (Sweden)

    Frances Dalton Cleaver

    2015-03-01

    Full Text Available This special issue furthers the study of natural resource management from a critical institutional perspective. Critical institutionalism (CI is a contemporary body of thought that explores how institutions dynamically mediate relationships between people, natural resources and society. It focuses on the complexity of institutions entwined in everyday social life, their historical formation, the interplay between formal and informal, traditional and modern arrangements, and the power relations that animate them. In such perspectives a social justice lens is often used to scrutinise the outcomes of institutional processes. We argue here that critical institutional approaches have potentially much to offer commons scholarship, particularly through the explanatory power of the concept of bricolage for better understanding institutional change.  Critical institutional approaches, gathering momentum over the past 15 years or so, have excited considerable interest but the insights generated from different disciplinary perspectives remain insufficiently synthesised. Analyses emphasising complexity can be relatively illegible to policy-makers, a fact which lessens their reach. This special issue therefore aims to synthesise critical institutional ideas and so to lay the foundation for moving beyond the emergent stage to make meaningful academic and policy impact. In bringing together papers here we define and synthesise key themes of critical institutionalism, outline the concept of institutional bricolage and identity some key challenges facing this school of thought.

  10. Critical Thinking as a Cognitive Educational Technology

    Directory of Open Access Journals (Sweden)

    Brylina Irina V.

    2016-01-01

    Full Text Available The article deals with higher education issues related to the formation of students’ intellectual work skills. The research objective of the paper was to consider critical thinking as a cognitive technology in education. In this regard, the didactic and structural approaches to the study of critical thinking do not contradict one another: each approach is a logical complement of the other and reveals certain aspects of the complex concept of critical thinking, giving emphasis to the argument, which is a tool, used both in critical and dogmatic thinking. By the general competence we mean principles of thinking, the ability to produce a reasoned piece of oral and written language, understanding and analysis of philosophical issues, considering the essence and value of the information. Among the professional competencies, the following should be listed: the ability to reconsider the gathered experience critically, the ability to collect, process, and interpret the data of modern research, to form judgments about the value and impact of the professional activity. The logical competence draws focused attention to the critical argument, regarding it throughout the course Logic. It is concluded that critical thinking can be seen as a cognitive educational technology for the formation of logical competence.

  11. Erik Erikson: critical times, critical theory.

    Science.gov (United States)

    Douvan, E

    1997-01-01

    The work and legacy of Erik Erikson are described in this brief outline of his career, his theories, and his impact on psychoanalysis, psychology, history, and the broader culture. His conception of the adolescent task-weaving internal tastes, talents, and values together with elements of one's life history and the demands of one's culture into a coherent identity-has had profound effects on developmental psychology and the way in which sophisticated youth construct and describe their lives. His extension of development through adulthood and old age established the field of life course development. His emphasis on the impact of history and culture on development was a critical element in the developing field of ego psychology. Many of his major contributions can be fruitfully understood in the context of his personal history and individual qualities.

  12. Critical behaviors near the (tri-)critical end point of QCD within the NJL model

    CERN Document Server

    Lu, Ya; Cui, Zhu-Fang; Zong, Hong-Shi

    2015-01-01

    We investigate the dynamical chiral symmetry breaking and its restoration at finite density and temperature within the two-flavor Nambu-Jona-Lasinio model, and mainly focus on the critical behaviors near the critical end point (CEP) and tricritical point (TCP) of QCD. The co-existence region of the Wigner and Nambu phase is determined in the phase diagram for the massive and massless current quark, respectively. We use the various susceptibilities to locate the CEP/TCP and then extract the critical exponents near them. Our calculations reveal that the various susceptibilities share the same critical behaviors for the physical current quark mass, while they show different features in the chiral limit. Furthermore the critical exponent of order parameter at the TCP, $\\beta$=1/4, differs from that on the $O(4)$ line, $\\beta$=1/2, which indicates a change in the universality class.

  13. Critical Management in Knowledge Organizations

    Science.gov (United States)

    Macpherson, Reynold

    2008-01-01

    Purpose: The purpose of this paper is to invite educational managers and management educators to reflect critically on practice. Design/methodology/approach: Using the point of Socrates' death, the paper suggests ways of reflecting on actions using ethically-critical, socially-critical, environmentally-critical, politically-critical and…

  14. The Anatomy of Critical Discourse.

    Science.gov (United States)

    Rosenfield, Lawrence W.

    1968-01-01

    Critical discourse is best understood when its logical features are identified. An examination of the basic elements and modes of rhetorical criticism (a form of critical discourse) produces a finite set of options for the critic, thus enabling him to develop a system of alternatives in his critical efforts. For example, by selecting from among…

  15. Criticality in Plasma Membranes

    Science.gov (United States)

    Machta, Benjamin; Papanikolaou, Stefanos; Sethna, James; Veatch, Sarah

    2011-03-01

    We are motivated by recent observations of micron-sized critical fluctuations in the 2d Ising Universality class in plasma membrane vesicles that are isolated from cortical cytoskeleton. We construct a minimal model of the plasma membrane's interaction with intact cytoskeleton which explains why large scale phase separation has not been observed in Vivo. In addition, we use analytical techniques from conformal field theory and numerical simulations to investigate the form of effective forces mediated by the membrane's proximity to criticality. We show that the range of this force is maximized near a critical point and we quantify its usefulness in mediating communication using techniques from information theory. Finally we use theoretical techniques from statistical physics in conjunction with Monte-Carlo simulations to understand how criticality can be used to increase the efficiency of membrane bound receptor mediated signaling. We expect that this sort of analysis will be broadly useful in understanding and quantifying the role of lipid ``rafts'' in a wide variety of membrane bound processes. Generally, we demonstrate that critical fluctuations provide a physical mechanism to organize and spatially segregate membrane components by providing channels for interaction over relatively large distances.

  16. Critical Environmental Regions

    Directory of Open Access Journals (Sweden)

    VICTOR SOROCOVSCHI

    2005-01-01

    Full Text Available A short etymological interpretation of the notion of regions (Rette Lineatte, etc.. The region is: R= f (S+P, where S is space and P is power. There follows an evaluation of the characteristics of the region and the presentation of different approaches to the region. From the classic ideas (von Humboldt, 1885, Dokuceaev, 1899, Herbertson, 1905, and others we get to a wide interpretative array of what we accept as organizational spatial units of geographical reality. The environmental region has important connotations with regard to the system as a surrounded element (man, society and the adjacent system. Critical environmental regions are areas where there already exists interactive degradation. The critical character may be physical, hence the “geocritical regions” or the result of human impact, hence the “anthropocritical regions.” Critical situations are differentiated at the local, regional, and global level. In order to understand critical regional situations we must refer to the following characteristics: fragility, resilience, and vulnerability. Still there are few environmental studies on critical regions and work must be done in this field.

  17. Stepping out of the Academic Brew: Using Critical Research to Break Down Hierarchies of Knowledge Production

    Science.gov (United States)

    Kress, Tricia M.

    2011-01-01

    Critical theory and critical research are undeniably useful for revealing oppressive social structures and challenging the status quo in the realm of grand theory; yet, they are also useful for creating knowledge structures when academics deploy them on the ground. This article explores how critical theory and critical research can be used to…

  18. Critical Narrative Inquiry

    DEFF Research Database (Denmark)

    While organizations have become central for thinking and structuring contemporary social action, existing perspectives on what they are and how to deal with them are still rooted in modern ideas about the foundations of society. The chapters in this volume take critical narrative inquiry — inspired...... by postmodern or post-human approaches to organizations — as a broad range of research and development strategies that challenge the dominant perspectives prevalent in the organizational literature. The purpose of the volume is three-fold. Firstly, a critical reading of organizations foregrounding notions...... of power and ethics is presented. Secondly, a new framework for understanding and analyzing organizational action based on critical notions of storytelling and sustainability is unfolded. Thirdly, the framework is deployed through innovative concepts and learning methodologies for leadership...

  19. Apoptosis in Critical Conditions

    Directory of Open Access Journals (Sweden)

    A. M. Golubev

    2006-01-01

    Full Text Available Apoptosis is a variant of programmed cell death. This term was introduced by Kerr et al. in 1972, but information on the important role of apoptosis of some cells in critical conditions has recently appeared. The review of literature considers the basic mechanisms of induction, development, and regulation of apoptosis. Based on a literature update, the authors analyze the role of apoptosis in the pathogenesis of various critical conditions: acute lung lesion (neutrophilic and epithelial hypotheses, sepsis, myocardial infarction, and ischemic stroke (apoptosis of tubular epithelial cells, hepatic dysfunction in sepsis, myopathies in critical conditions. The data of studies dealing with the effects of inhaled and non-inhaled anesthetics on the apoptosis of neurons of the brain and lymphocytes are given. The review of literature presents the options of therapeutic apoptosis modulation by pharmacological methods.  

  20. Critical Axial Load

    Directory of Open Access Journals (Sweden)

    Walt Wells

    2008-01-01

    Full Text Available Our objective in this paper is to solve a second order differential equation for a long, simply supported column member subjected to a lateral axial load using Heun's numerical method. We will use the solution to find the critical load at which the column member will fail due to buckling. We will calculate this load using Euler's derived analytical approach for an exact solution, as well as Euler's Numerical Method. We will then compare the three calculated values to see how much they deviate from one another. During the critical load calculation, it will be necessary to calculate the moment of inertia for the column member.

  1. The critical interface

    DEFF Research Database (Denmark)

    Pold, Søren

    2005-01-01

    In this paper, I argue that aesthetics might serve the role of critical reflection on representation within the field of computing, and I demonstrate through three analytic examples from computer art how this might look like and what the contribution might be.......In this paper, I argue that aesthetics might serve the role of critical reflection on representation within the field of computing, and I demonstrate through three analytic examples from computer art how this might look like and what the contribution might be....

  2. Critical Narrative Inquiry

    DEFF Research Database (Denmark)

    by postmodern or post-human approaches to organizations — as a broad range of research and development strategies that challenge the dominant perspectives prevalent in the organizational literature. The purpose of the volume is three-fold. Firstly, a critical reading of organizations foregrounding notions...... of power and ethics is presented. Secondly, a new framework for understanding and analyzing organizational action based on critical notions of storytelling and sustainability is unfolded. Thirdly, the framework is deployed through innovative concepts and learning methodologies for leadership...

  3. Critical care transport.

    Science.gov (United States)

    Williams, Kenneth A; Sullivan, Francis M

    2013-12-03

    Critical care transport (CCT) is the segment of the Emergency Medical Services (EMS) system that transports patients who are critically ill or injured. Nearly 1,000 medical helicopters affiliated with over 300 transport programs, hundreds of fixed-wing aircraft, and many, many ground ambulances assisting adult, pediatric and neonatal CCT teams are operating in the United States.1 This article reviews the history of and indications for CCT, team qualifications, vehicle options, safety, CCT system design, and physician involvement in CCT. It concludes with a brief review of CCT services in Rhode Island.

  4. Critical Pedagogy Principles in Teaching EFL Reading

    Directory of Open Access Journals (Sweden)

    Slamet Wahyudi Yulianto

    2015-12-01

    Full Text Available This study attempts to reveal how the use of critical pedagogy principles in teaching English as a Foreign Language (EFL reading facilitates students to think critically. Additionally, it investigates and elaborates the benefits and challenges of using critical pedagogy principles in teaching EFL reading. The three critical pedagogy principles used in this study were dialogic education, democratic classroom, and reading the world and the word. Critical thinking skills and dispositions expected to be performed by the participants were analysis and evaluation skills, open-mindedness, and making reasoned decision. This is a case study design which was conducted in the form of teaching program. The teaching program which consisted of eight meetings was given to 59 EFL sophomores in the Reading in Professional Context class at a private teacher education in Bandung. Data in the form of classroom talks and activities and students‟ responses as well as their critical thinking skills self-assessment were collected by using video recordings, observation notes, interview guideline, students‟ learning journals, and questionnaires. It is revealed that the teaching program has facilitated students to think critically by providing four categories of activity. They are (1 offering problematic topics and reading materials that are linked to the students‟ lives, (2 encouraging students to read between the lines, (3 distributing classroom power, and (4 creating space for students‟ voices to be heard. Meanwhile, there are two benefits of the teaching program, namely (1 language development and (2 new knowledge as well as experience acquisition. However, there are three major challenges in conducting the teaching program that are (1 the lack of classroom-friendly authentic controversial reading materials, (2 the passive culture, and (3 the unpredictable classroom.

  5. Using the Social Studies Textbook to Teach Critical Thinking.

    Science.gov (United States)

    Wright, Ian

    1996-01-01

    Considers the ubiquitous use of textbooks in social studies classrooms and recommends constructing critical-thinking exercises revealing textual bias. Suggested exercises concern issues of emotional or ambiguous language, geographic and cultural inclusion/exclusion, and accuracy of data. Includes several examples of critical-thinking activities.…

  6. Critical Thinking in Social Work Education: A Research Synthesis

    Science.gov (United States)

    Samson, Patricia L.

    2016-01-01

    In a meta-analytic review of critical thinking in social work education, findings revealed variability in research designs, methods, and subsequent findings. The 10 studies reviewed assessed different components of critical thinking and highlighted different potential moderator variables. Although there are significant limitations to all the…

  7. A critical systems perspective on the design of organizational space

    NARCIS (Netherlands)

    Mobach, Mark P.

    2007-01-01

    This paper is the first to introduce critical systems thinking into a new emerging research strand: the design of organizational space. The study revealed two things. First, critical systems thinking provides a thorough framework to understand the possibilities to connect organization and building;

  8. TypeScript revealed

    CERN Document Server

    Maharry, Dan

    2013-01-01

    TypeScript Revealed is a quick 100-page guide to Anders Hejlsberg's new take on JavaScript. With this brief, fast-paced introduction to TypeScript, .NET, Web and Windows 8 application developers who are already familiar with JavaScript will easily get up to speed with TypeScript and decide whether or not to start incorporating it into their own development. TypeScript is 'JavaScript for Application-scale development'; a superset of JavaScript that brings to it an additional object-oriented-like syntax familiar to .NET programmers that compiles down into simple, clean JavaScript that any browse

  9. Revealing the programming process

    DEFF Research Database (Denmark)

    Bennedsen, Jens; Caspersen, Michael Edelgaard

    2005-01-01

    One of the most important goals of an introductory programming course is that the students learn a systematic approach to the development of computer programs. Revealing the programming process is an important part of this; however, textbooks do not address the issue -- probably because the textb......One of the most important goals of an introductory programming course is that the students learn a systematic approach to the development of computer programs. Revealing the programming process is an important part of this; however, textbooks do not address the issue -- probably because...... the textbook medium is static and therefore ill-suited to expose the process of programming. We have found that process recordings in the form of captured narrated programming sessions are a simple, cheap, and efficient way of providing the revelation.We identify seven different elements of the programming...... process for which process recordings are a valuable communication media in order to enhance the learning process. Student feedback indicates both high learning outcome and superior learning potential compared to traditional classroom teaching....

  10. Translation as Literary Criticism.

    Science.gov (United States)

    di Stefano, B. Follkart

    1982-01-01

    It is proposed that literary translation is intrinsically an act of literary criticism. This theory is illustrated by discussion of specific problems in translating Sartre's "La Nausee" and Leonard Forest's "Le pays de la Sagouine," especially the use of verb tense. (MSE)

  11. Critical-Illness Polyneuropathy

    OpenAIRE

    J Gordon Millichap

    1993-01-01

    A review of the neurological complications of sepsis from the University of Western Ontario, London, and Queen’s University, Kingston, Ontario, Canada, draws attention to “critical-illness polyneuropathy” as a cause of difficulty in weaning from the ventilator.

  12. Reply to critics

    NARCIS (Netherlands)

    Lindahl, H.K.

    2014-01-01

    In my reply to critics I address a raft of issues raised by the commentators to Fault Lines of Globalization. These issues include: the radicalization of the Husserlian concept of intentionality as differance; the irreducibility of a-legality to (il)legality; the relation between legal orders and

  13. Views on Critical Pedagogy

    Institute of Scientific and Technical Information of China (English)

    李怡静

    2015-01-01

    Instead of being a theory Critical Pedagogy is a way of teaching. It tries to establish a bridge between the classroom and the wider social context, and mainly deals with power, such as social justice and social change through education. Based on the idea that education should be equal to everyone, CP never stops exploring the right way of realizing education equality.

  14. Translation as Literary Criticism.

    Science.gov (United States)

    di Stefano, B. Follkart

    1982-01-01

    It is proposed that literary translation is intrinsically an act of literary criticism. This theory is illustrated by discussion of specific problems in translating Sartre's "La Nausee" and Leonard Forest's "Le pays de la Sagouine," especially the use of verb tense. (MSE)

  15. Introducing Veteran Critical Theory

    Science.gov (United States)

    Phillips, Glenn A.; Lincoln, Yvonna S.

    2017-01-01

    Research on student veterans is in an infant state. As veterans continue to enroll in institutions of higher education, researchers must explore new ways of "knowing" student veterans. It is not enough to only describe and model this growing demographic, researchers must also have a tool for criticism and question. The next in an…

  16. TQM in critical care.

    Science.gov (United States)

    Massarweh, L J

    1998-06-01

    No consistently accepted methods reliably measure quality differences among and within critical care units (CCUs). With total quality management, physicians and nurses have the framework to accurately assess the organizational climate of their CCU. A study asks nurses to identify organizational components in their CCU.

  17. Reply to critics

    NARCIS (Netherlands)

    Lindahl, H.K.

    2014-01-01

    In my reply to critics I address a raft of issues raised by the commentators to Fault Lines of Globalization. These issues include: the radicalization of the Husserlian concept of intentionality as differance; the irreducibility of a-legality to (il)legality; the relation between legal orders and so

  18. Critical Points of Contact

    DEFF Research Database (Denmark)

    Jensen, Ole B.; Morelli, Nicola

    2011-01-01

    where the networks meet and establish contact. Thus we argue for the usefulness of the notion of Critical Point of Contact (CPC) to deepen our understanding of the actual life within networks. En route to this notion we draw upon theories within as diverse realms such as interaction design, service...

  19. Critical Points of Contact

    DEFF Research Database (Denmark)

    2012-01-01

    In this brief article, we shall illustrate the application of the analytical and interventionist concept of ‘Critical Points of Contact’ (CPC) through a number of urban design studios. The notion of CPC has been developed over a span of the last three to four years and is reported in more detail...

  20. APS Quantum Critical Higgs

    CERN Document Server

    Bellazzini, Brando; Hubisz, Jay; Lee, Seung J.; Serra, Javi; Terning, John

    2016-01-01

    The appearance of the light Higgs boson at the LHC is difficult to explain, particularly in light of naturalness arguments in quantum field theory. However light scalars can appear in condensed matter systems when parameters (like the amount of doping) are tuned to a critical point. At zero temperature these quantum critical points are directly analogous to the finely tuned standard model. In this paper we explore a class of models with a Higgs near a quantum critical point that exhibits non-mean-field behavior. We discuss the parametrization of the effects of a Higgs emerging from such a critical point in terms of form factors, and present two simple realistic scenarios based on either generalized free fields or a 5D dual in AdS space. For both of these models we consider the processes $gg\\to ZZ$ and $gg\\to hh$, which can be used to gain information about the Higgs scaling dimension and IR transition scale from the experimental data.

  1. Infancy and Critical Theory

    OpenAIRE

    UENO, Toshiya

    2015-01-01

    This paper examines with a relationship between infancy and critical theory by addressing a Japanese postwar intellectual who is a kind of counterparts of Frankfurt School scholar in Euro/Am context, Fujita Shozo, and one of his texts in the details.

  2. Replies to Criticisms

    Science.gov (United States)

    Hamilton, James R.

    2009-01-01

    In this essay, Hamilton responds to criticisms of his book "The Art of Theater" (Malden, MA: Blackwell, 2007). Acknowledging that he expected that the central proposals in "The Art of Theater" would seem a little strange to philosophers, he reiterates his belief that the three general facts of any theatrical performance are its presentation, its…

  3. The critical interface

    DEFF Research Database (Denmark)

    Pold, Søren

    2005-01-01

    In this paper, I argue that aesthetics might serve the role of critical reflection on representation within the field of computing, and I demonstrate through three analytic examples from computer art how this might look like and what the contribution might be....

  4. Situation criticality and basketball officials' stress levels.

    Science.gov (United States)

    Ritchie, Jason; Basevitch, Itay; Rodenberg, Ryan; Tenenbaum, Gershon

    2017-11-01

    Officials are expected to perform impeccably despite the wide range of stressors they experience. A stressor that officials frequently report is situation criticality. Situation criticality is comprised of score differential (i.e., more pressure in close games) and time remaining in a game (i.e., more pressure as time expires), which affects athletes' stress levels. The present study explored the effect of situation criticality on officials' stress levels. High school basketball officials (n = 108) with an average of 18.1 (SD = 11.2) years of officiating experience were given a survey packet containing game situations that varied in criticality. For each game situation (n = 9) officials completed the overall stress and appraisal portions of the Stress Appraisal Measure (SAM). Results revealed that situation criticality has an effect on officials' perceived stress levels. Both threat and challenge appraisals were positively correlated with perceived stress. Overall, these findings indicate that officials' stress levels fluctuate within games depending on score differential and time of game. The findings encourage officials to recognise and manage their stress, possibly through their appraisals. Additionally, the findings can affect the training of officials in the management of stress, as well as prompt the consideration of potential rule changes that reflect the increased situational demands on officials in critical situations (e.g., expanded instant replay).

  5. Controlling superconductivity by tunable quantum critical points.

    Science.gov (United States)

    Seo, S; Park, E; Bauer, E D; Ronning, F; Kim, J N; Shim, J-H; Thompson, J D; Park, Tuson

    2015-03-04

    The heavy fermion compound CeRhIn5 is a rare example where a quantum critical point, hidden by a dome of superconductivity, has been explicitly revealed and found to have a local nature. The lack of additional examples of local types of quantum critical points associated with superconductivity, however, has made it difficult to unravel the role of quantum fluctuations in forming Cooper pairs. Here, we show the precise control of superconductivity by tunable quantum critical points in CeRhIn5. Slight tin-substitution for indium in CeRhIn5 shifts its antiferromagnetic quantum critical point from 2.3 GPa to 1.3 GPa and induces a residual impurity scattering 300 times larger than that of pure CeRhIn5, which should be sufficient to preclude superconductivity. Nevertheless, superconductivity occurs at the quantum critical point of the tin-doped metal. These results underline that fluctuations from the antiferromagnetic quantum criticality promote unconventional superconductivity in CeRhIn5.

  6. Critical Education, Critical Pedagogies, Marxist Education in the United States

    Science.gov (United States)

    Foley, Jean Ann; Morris, Doug; Gounari, Panayota; Agostinone-Wilson, Faith

    2015-01-01

    As critical pedagogy becomes more mainstream on the educational landscape in the United States, it is important to revisit the original tenets of critical pedagogy and explore their current manifestations. Since the beginning of "criticalism" from the theoretical/foundational work of the Frankfurt School of Critical Social Theory,…

  7. Critical Education, Critical Pedagogies, Marxist Education in the United States

    Science.gov (United States)

    Foley, Jean Ann; Morris, Doug; Gounari, Panayota; Agostinone-Wilson, Faith

    2015-01-01

    As critical pedagogy becomes more mainstream on the educational landscape in the United States, it is important to revisit the original tenets of critical pedagogy and explore their current manifestations. Since the beginning of "criticalism" from the theoretical/foundational work of the Frankfurt School of Critical Social Theory,…

  8. Android Emotions Revealed

    DEFF Research Database (Denmark)

    Vlachos, Evgenios; Schärfe, Henrik

    2012-01-01

    This work presents a method for designing facial interfaces for sociable android robots with respect to the fundamental rules of human affect expression. Extending the work of Paul Ekman towards a robotic direction, we follow the judgment-based approach for evaluating facial expressions to test...... in which case an android robot like the Geminoid|DK –a duplicate of an Original person- reveals emotions convincingly; when following an empirical perspective, or when following a theoretical one. The methodology includes the processes of acquiring the empirical data, and gathering feedback on them. Our...... findings are based on the results derived from a number of judgments, and suggest that before programming the facial expressions of a Geminoid, the Original should pass through the proposed procedure. According to our recommendations, the facial expressions of an android should be tested by judges, even...

  9. Android Emotions Revealed

    DEFF Research Database (Denmark)

    Vlachos, Evgenios; Schärfe, Henrik

    2012-01-01

    This work presents a method for designing facial interfaces for sociable android robots with respect to the fundamental rules of human affect expression. Extending the work of Paul Ekman towards a robotic direction, we follow the judgment-based approach for evaluating facial expressions to test...... in which case an android robot like the Geminoid|DK –a duplicate of an Original person- reveals emotions convincingly; when following an empirical perspective, or when following a theoretical one. The methodology includes the processes of acquiring the empirical data, and gathering feedback on them. Our...... findings are based on the results derived from a number of judgments, and suggest that before programming the facial expressions of a Geminoid, the Original should pass through the proposed procedure. According to our recommendations, the facial expressions of an android should be tested by judges, even...

  10. Finding critical points whose polarization is also a critical point

    OpenAIRE

    Squassina, Marco; Van Schaftingen, Jean

    2011-01-01

    We show that near any given minimizing sequence of paths for the mountain pass lemma, there exists a critical point whose polarization is also a critical point. This is motivated by the fact that if any polarization of a critical point is also a critical point and the Euler-Lagrange equation is a second-order semi-linear elliptic problem, T. Bartsch, T. Weth and M. Willem (J. Anal. Math., 2005) have proved that the critical point is axially symmetric.

  11. Critical coupling in plasmonic resonator arrays

    Science.gov (United States)

    Balci, Sinan; Kocabas, Coskun; Aydinli, Atilla

    2011-08-01

    We report critical coupling of electromagnetic waves to plasmonic cavity arrays fabricated on Moiré surfaces. Dark field plasmon microscopy imaging and polarization dependent spectroscopic reflection measurements reveal the critical coupling conditions of the cavities. The critical coupling conditions depend on the superperiod of the Moiré surface, which also defines the coupling between the cavities. Complete transfer of the incident power can be achieved for traveling wave plasmonic resonators, which have a relatively short superperiod. When the superperiod of the resonators increases, the coupled resonators become isolated standing wave resonators in which complete transfer of the incident power is not possible. Analytical and finite difference time domain calculations support the experimental observations.

  12. Shape phase mixing in critical point nuclei

    CERN Document Server

    Budaca, R

    2016-01-01

    Spectral properties of nuclei near the critical point of the quantum phase transition between spherical and axially symmetric shapes are studied in a hybrid collective model which combines the $\\gamma$-stable and $\\gamma$-rigid collective conditions through a rigidity parameter. The model in the lower and upper limits of the rigidity parameter recovers the X(5) and X(3) solutions respectively, while in the equally mixed case it corresponds to the X(4) critical point symmetry. Numerical applications of the model on nuclei from regions known for critical behavior reveal a sizable shape phase mixing and its evolution with neutron or proton numbers. The model also enables a better description of energy spectra and electromagnetic transitions for these nuclei.

  13. Critical Landau velocity in helium nanodroplets.

    Science.gov (United States)

    Brauer, Nils B; Smolarek, Szymon; Loginov, Evgeniy; Mateo, David; Hernando, Alberto; Pi, Marti; Barranco, Manuel; Buma, Wybren J; Drabbels, Marcel

    2013-10-11

    The best-known property of superfluid helium is the vanishing viscosity that objects experience while moving through the liquid with speeds below the so-called critical Landau velocity. This critical velocity is generally considered a macroscopic property as it is related to the collective excitations of the helium atoms in the liquid. In the present work we determine to what extent this concept can still be applied to nanometer-scale, finite size helium systems. To this end, atoms and molecules embedded in helium nanodroplets of various sizes are accelerated out of the droplets by means of optical excitation, and the speed distributions of the ejected particles are determined. The measurements reveal the existence of a critical velocity in these systems, even for nanodroplets consisting of only a thousand helium atoms. Accompanying theoretical simulations based on a time-dependent density functional description of the helium confirm and further elucidate this experimental finding.

  14. Algorithms of wave reflective critical angle on interface

    Science.gov (United States)

    Zhang, YongGang; Zhang, JianXue; Jiao, Lin; Li, Qinghong

    2017-02-01

    This article is in connection with calculating of reflection critical angle on interface, author has found that reflective wave have a quarter wavelength effects and deduced both absolute and relative reflection critical Angle calculation formulas. The two formulas can easy solve the question of reflection critical angle on interface of one side of the air where it is not calculated by Snell's law. Snell's law only reveals that rate of the wave velocity projected to the interface, the methods of this paper reveal the normal component of wave velocity on the interface relationship. The methods will be widely used in various fields such as light, electromagnetic waves, sound waves and water waves etc.

  15. Human immunodeficiency virus impairs reverse cholesterol transport from macrophages.

    Directory of Open Access Journals (Sweden)

    Zahedi Mujawar

    2006-10-01

    Full Text Available Several steps of HIV-1 replication critically depend on cholesterol. HIV infection is associated with profound changes in lipid and lipoprotein metabolism and an increased risk of coronary artery disease. Whereas numerous studies have investigated the role of anti-HIV drugs in lipodystrophy and dyslipidemia, the effects of HIV infection on cellular cholesterol metabolism remain uncharacterized. Here, we demonstrate that HIV-1 impairs ATP-binding cassette transporter A1 (ABCA1-dependent cholesterol efflux from human macrophages, a condition previously shown to be highly atherogenic. In HIV-1-infected cells, this effect was mediated by Nef. Transfection of murine macrophages with Nef impaired cholesterol efflux from these cells. At least two mechanisms were found to be responsible for this phenomenon: first, HIV infection and transfection with Nef induced post-transcriptional down-regulation of ABCA1; and second, Nef caused redistribution of ABCA1 to the plasma membrane and inhibited internalization of apolipoprotein A-I. Binding of Nef to ABCA1 was required for down-regulation and redistribution of ABCA1. HIV-infected and Nef-transfected macrophages accumulated substantial amounts of lipids, thus resembling foam cells. The contribution of HIV-infected macrophages to the pathogenesis of atherosclerosis was supported by the presence of HIV-positive foam cells in atherosclerotic plaques of HIV-infected patients. Stimulation of cholesterol efflux from macrophages significantly reduced infectivity of the virions produced by these cells, and this effect correlated with a decreased amount of virion-associated cholesterol, suggesting that impairment of cholesterol efflux is essential to ensure proper cholesterol content in nascent HIV particles. These results reveal a previously unrecognized dysregulation of intracellular lipid metabolism in HIV-infected macrophages and identify Nef and ABCA1 as the key players responsible for this effect. Our findings

  16. Society of Critical Care Medicine

    Science.gov (United States)

    ... Critical Care Medicine Podcasts Hosts iCritical Care App Social Media Critical Care Statistics eCommunity Media Relations SmartBrief SCCM App Education Center Annual Congress Program Abstracts Registration and Hotel Exhibits-Sponsorship Sightseeing Activities Past and Future Critical ...

  17. Critical Analysis of Multimodal Discourse

    DEFF Research Database (Denmark)

    van Leeuwen, Theo

    2013-01-01

    This is an encyclopaedia article which defines the fields of critical discourse analysis and multimodality studies, argues that within critical discourse analysis more attention should be paid to multimodality, and within multimodality to critical analysis, and ends reviewing a few examples...... of recent work in the critical analysis of multimodal discourse....

  18. Affecting Critical Thinking through Speech.

    Science.gov (United States)

    O'Keefe, Virginia P.

    Intended for teachers, this booklet shows how spoken language can affect student thinking and presents strategies for teaching critical thinking skills. The first section discusses the theoretical and research bases for promoting critical thinking through speech, defines critical thinking, explores critical thinking as abstract thinking, and tells…

  19. Toward a Gramscian Critical Rhetoric.

    Science.gov (United States)

    Zompetti, Joseph P.

    1997-01-01

    Contends A. Gramsci can provide a perspective on the cultural dominance of rhetoric and formation of a critical "telos"--his work can contribute to understanding critical rhetoric. Demonstrates that Gramscian notions can extend critical rhetoric into an enterprise that permits critical self-reflexivity and praxis and create new…

  20. Critical branching neural networks.

    Science.gov (United States)

    Kello, Christopher T

    2013-01-01

    It is now well-established that intrinsic variations in human neural and behavioral activity tend to exhibit scaling laws in their fluctuations and distributions. The meaning of these scaling laws is an ongoing matter of debate between isolable causes versus pervasive causes. A spiking neural network model is presented that self-tunes to critical branching and, in doing so, simulates observed scaling laws as pervasive to neural and behavioral activity. These scaling laws are related to neural and cognitive functions, in that critical branching is shown to yield spiking activity with maximal memory and encoding capacities when analyzed using reservoir computing techniques. The model is also shown to account for findings of pervasive 1/f scaling in speech and cued response behaviors that are difficult to explain by isolable causes. Issues and questions raised by the model and its results are discussed from the perspectives of physics, neuroscience, computer and information sciences, and psychological and cognitive sciences.

  1. Critical Digital Tourism Studies

    DEFF Research Database (Denmark)

    Gyimóthy, Szilvia; Munar, Ana María

    2013-01-01

    ) are approached mostly from a business administration perspective and informed by conceptual frameworks developed in management and marketing. IT studies in tourism are still at a stage similar to the first advocacy phase of tourism research in general (Jafari, 1990) and are seldom inspired by relevant......This paper advocates the need for a critical and cross-disciplinary research agenda on the field of digital technologies and tourism. The changing virtual landscape of tourism has received increased attention by tourism scholars. However, contemporary studies on information technologies (IT...... to studying digital socio-technical systems and virtual mediation in tourism. Critical Digital Tourism Studies opens a new cross-disciplinary field where the sociality of virtual tourism interactions is examined (entailing the study of structures, social rules, ideologies, power relations, sustainability...

  2. Ghosts of Critical Gravity

    CERN Document Server

    Porrati, Massimo

    2011-01-01

    Recently proposed "critical" higher-derivative gravities in $AdS_D$ $D>3$ are expected to carry logarithmic representation of the Anti de Sitter isometry group. In this note, we quantize linear fluctuations of these critical gravities, which are known to be either identical with linear fluctuations of Einstein's gravity or else satisfy logarithmic boundary conditions at spacial infinity. We identify the scalar product uniquely defined by the symplectic structure implied by the classical action, and show that it does not posses null vectors. Instead, we show that the scalar product between any two Einstein modes vanishes, while the scalar product of an Einstein mode with a logarithmic mode is generically nonzero. This is the basic property of logarithmic representation that makes them neither unitary nor unitarizable.

  3. Drug related critical incidents.

    Science.gov (United States)

    Khan, F A; Hoda, M Q

    2005-01-01

    Drug related incidents are a common form of reported medical errors. This paper reviews the critical incidents related to drug errors reported from the main operating theatre suite in a teaching hospital in a developing country from January 1997 to December 2002. Each report was evaluated individually by two reviewers using a structured process. During this period, 44 874 anaesthetics were administered; 768 critical incidents were reported, 165 (21%) of which were related to drug errors. Underdosage, side-effect/drug reaction and syringe swap were the most common. A total of 76% were classified as preventable; 56% due to human error and 19% due to system error. High risk incidents accounted for 10% of all drug errors and most of these were related to the use of neuromuscular blocking drugs. This analysis has been found useful in addressing some issues about priorities.

  4. Critical Digital Tourism Studies

    DEFF Research Database (Denmark)

    Gyimóthy, Szilvia; Munar, Ana María

    2013-01-01

    ) are approached mostly from a business administration perspective and informed by conceptual frameworks developed in management and marketing. IT studies in tourism are still at a stage similar to the first advocacy phase of tourism research in general (Jafari, 1990) and are seldom inspired by relevant......This paper advocates the need for a critical and cross-disciplinary research agenda on the field of digital technologies and tourism. The changing virtual landscape of tourism has received increased attention by tourism scholars. However, contemporary studies on information technologies (IT...... to studying digital socio-technical systems and virtual mediation in tourism. Critical Digital Tourism Studies opens a new cross-disciplinary field where the sociality of virtual tourism interactions is examined (entailing the study of structures, social rules, ideologies, power relations, sustainability...

  5. Critical Mandelbrot Cascades

    Science.gov (United States)

    Barral, Julien; Kupiainen, Antti; Nikula, Miika; Saksman, Eero; Webb, Christian

    2014-01-01

    We study Mandelbrot's multiplicative cascade measures at the critical temperature. As has been recently shown by Barral et al. (C R Acad Sci Paris Ser I 350:535-538, 2012), an appropriately normalized sequence of cascade measures converges weakly in probability to a nontrivial limit measure. We prove that these limit measures have no atoms and give bounds for the modulus of continuity of the cumulative distribution function of the measure. Using the earlier work of Barral and Seuret (Adv Math 214:437-468, 2007), we compute the multifractal spectrum of the measures. We also extend the result of Benjamini and Schramm (Commun Math Phys 289:653-662, 2009), in which the KPZ formula from quantum gravity is validated for the high temperature cascade measures, to the critical and low temperature cases.

  6. Criticality safety training

    Energy Technology Data Exchange (ETDEWEB)

    Woodruff, S.K. [Los Alamos National Lab., NM (United States)

    1997-06-01

    Criticality safety training is an important element of the Plutonium Facility safety program at Los Alamos National Laboratory. Training consists of student self-study handbooks and hands-on performance-based training in a mock-up laboratory containing gloveboxes, trolley conveyor system, and self-monitoring instruments. A 10-minute video tape and lecture was presented to describe how training in this area is conducted.

  7. Critical care cardiology.

    Science.gov (United States)

    Marks, S L; Abbott, J A

    1998-11-01

    Emergency management of the patient with cardiac disease is an important part of veterinary practice. Although the causes of cardiac disease may be diverse, the understanding of basic pathophysiology will enable the clinician to formulate a rational diagnostic and therapeutic plan. The veterinary clinician must be able to triage the emergency patient, assess the clinical condition, and provide appropriate therapy. Close monitoring of the critically ill patient is crucial to patient survival and will help tailor therapy.

  8. Criticisms of Relevance Theory

    Institute of Scientific and Technical Information of China (English)

    尚静; 孟晔; 焦丽芳

    2006-01-01

    This paper briefly introduces first the notion of Sperber and Wilson's Relevance Theory. Then, the motivation of S & W putting forward their RT is also mentioned. Secondly, the paper gives some details about the methodology of RT, in which ostensive-inferential communication, context and optimal relevance are highlighted. Thirdly, the paper focuses on the criticisms of RT from different areas of research on human language and communication. Finally, the paper draws a conclusion on the great importance of RT in pragmatics.

  9. Carahunge - A Critical Assessment

    Science.gov (United States)

    González-García, A. César

    Carahunge is a megalithic monument in southern Armenia that has often been acclaimed as the oldest observatory. The monument, composed of dozens of standing stones, has some perforated stones. The direction of the holes has been measured and their orientation is related to the sun, moon, and stars, obtaining a date for the construction of such devices. After a critical review of the methods and conclusions, these are shown as untenable.

  10. Education for Critical Thinking

    Science.gov (United States)

    2013-02-01

    develop critical thinkers because knowing the pit - falls of logic can hone one’s understanding. Because logic is so important for parsing complexity...provide answers. We are looking for a program similar to what the Naval War College famously did during the 1930s when leaders like William “ Bull ...the 1930s when leaders like William “ Bull ” Halsey not only ex- changed ideas but also had the chance to test “pet theories” in an uncon- strained

  11. Horizon as critical phenomenon

    Science.gov (United States)

    Lee, Sung-Sik

    2016-09-01

    We show that renormalization group flow can be viewed as a gradual wave function collapse, where a quantum state associated with the action of field theory evolves toward a final state that describes an IR fixed point. The process of collapse is described by the radial evolution in the dual holographic theory. If the theory is in the same phase as the assumed IR fixed point, the initial state is smoothly projected to the final state. If in a different phase, the initial state undergoes a phase transition which in turn gives rise to a horizon in the bulk geometry. We demonstrate the connection between critical behavior and horizon in an example, by deriving the bulk metrics that emerge in various phases of the U( N ) vector model in the large N limit based on the holographic dual constructed from quantum renormalization group. The gapped phase exhibits a geometry that smoothly ends at a finite proper distance in the radial direction. The geometric distance in the radial direction measures a complexity: the depth of renormalization group transformation that is needed to project the generally entangled UV state to a direct product state in the IR. For gapless states, entanglement persistently spreads out to larger length scales, and the initial state can not be projected to the direct product state. The obstruction to smooth projection at charge neutral point manifests itself as the long throat in the anti-de Sitter space. The Poincare horizon at infinity marks the critical point which exhibits a divergent length scale in the spread of entanglement. For the gapless states with non-zero chemical potential, the bulk space becomes the Lifshitz geometry with the dynamical critical exponent two. The identification of horizon as critical point may provide an explanation for the universality of horizon. We also discuss the structure of the bulk tensor network that emerges from the quantum renormalization group.

  12. Relative Critical Points

    Directory of Open Access Journals (Sweden)

    Debra Lewis

    2013-05-01

    Full Text Available Relative equilibria of Lagrangian and Hamiltonian systems with symmetry are critical points of appropriate scalar functions parametrized by the Lie algebra (or its dual of the symmetry group. Setting aside the structures – symplectic, Poisson, or variational – generating dynamical systems from such functions highlights the common features of their construction and analysis, and supports the construction of analogous functions in non-Hamiltonian settings. If the symmetry group is nonabelian, the functions are invariant only with respect to the isotropy subgroup of the given parameter value. Replacing the parametrized family of functions with a single function on the product manifold and extending the action using the (coadjoint action on the algebra or its dual yields a fully invariant function. An invariant map can be used to reverse the usual perspective: rather than selecting a parametrized family of functions and finding their critical points, conditions under which functions will be critical on specific orbits, typically distinguished by isotropy class, can be derived. This strategy is illustrated using several well-known mechanical systems – the Lagrange top, the double spherical pendulum, the free rigid body, and the Riemann ellipsoids – and generalizations of these systems.

  13. Relative Critical Points

    Science.gov (United States)

    Lewis, Debra

    2013-05-01

    Relative equilibria of Lagrangian and Hamiltonian systems with symmetry are critical points of appropriate scalar functions parametrized by the Lie algebra (or its dual) of the symmetry group. Setting aside the structures - symplectic, Poisson, or variational - generating dynamical systems from such functions highlights the common features of their construction and analysis, and supports the construction of analogous functions in non-Hamiltonian settings. If the symmetry group is nonabelian, the functions are invariant only with respect to the isotropy subgroup of the given parameter value. Replacing the parametrized family of functions with a single function on the product manifold and extending the action using the (co)adjoint action on the algebra or its dual yields a fully invariant function. An invariant map can be used to reverse the usual perspective: rather than selecting a parametrized family of functions and finding their critical points, conditions under which functions will be critical on specific orbits, typically distinguished by isotropy class, can be derived. This strategy is illustrated using several well-known mechanical systems - the Lagrange top, the double spherical pendulum, the free rigid body, and the Riemann ellipsoids - and generalizations of these systems.

  14. Diagnostics in critical conditions

    Directory of Open Access Journals (Sweden)

    SadchikovD.V.

    2011-09-01

    Full Text Available The purpose of research: improvement of quality of diagnostics at the patients in a critical condition in intensive care unit. Material and methods. In total have analyzed 1957 medical cards of the patients who have died in ICU»s. At the first stage studied the factors influencing on diagnostics of critically ill patients (medical cards of 1557 patients; at the second stage investigated influence of the diagnostic standards in ICU»s practice on improvement of quality of diag- nostics of critically ill patients (400 medical cards of the patients who have died. Entry criterions were standards and algorithm of diagnostics. Techniques of research: average bed-day in groups, first-day lethality, quantity of the carried out laboratory tests and tool methods of research, level of consciousness of the patients (Glasgow come score, severity of disease by ICU»s patients (APACHE II scores. Results. Quality of diagnostics depend on carried out laboratory tests and tool methods of research, level of consciousness of the patients (Glasgow come score, severity of disease by ICU»s patients (APACHE II score. The conclusion. The laboratory tests and tool methods of research conforming to the standards of diagnostics are necessary for improvement of quality of diagnostics, it is necessary to take into account an altered level of consciousness (Glasgow come score and severity of disease by ICU»s patients (APACHE II scores

  15. New Criticism and Poetry Interpretation

    Institute of Scientific and Technical Information of China (English)

    王洁成

    2012-01-01

      New Criticism is a dominant trend in literary criticism of the mid twentieth century. New Criticism, focusing on the text and emphasizing close reading, finds its excellence in poetry interpretation. This essay introduces the theoretical basis, the approach—close reading, and terms of New Criticism. The essay analyzes the metaphysical poem“Virtue”written by George Herbert by the use of New Criticism.

  16. Bioorthogonal mimetics of palmitoyl-CoA and myristoyl-CoA and their subsequent isolation by click chemistry and characterization by mass spectrometry reveal novel acylated host-proteins modified by HIV-1 infection.

    Science.gov (United States)

    Colquhoun, David R; Lyashkov, Alexey E; Ubaida Mohien, Ceereena; Aquino, Veronica N; Bullock, Brandon T; Dinglasan, Rhoel R; Agnew, Brian J; Graham, David R M

    2015-06-01

    Protein acylation plays a critical role in protein localization and function. Acylation is essential for human immunodeficiency virus 1 (HIV-1) assembly and budding of HIV-1 from the plasma membrane in lipid raft microdomains and is mediated by myristoylation of the Gag polyprotein and the copackaging of the envelope protein is facilitated by colocalization mediated by palmitoylation. Since the viral accessory protein NEF has been shown to alter the substrate specificity of myristoyl transferases, and alter cargo trafficking lipid rafts, we hypothesized that HIV-1 infection may alter protein acylation globally. To test this hypothesis, we labeled HIV-1 infected cells with biomimetics of acyl azides, which are incorporated in a manner analogous to natural acyl-Co-A. A terminal azide group allowed us to use a copper catalyzed click chemistry to conjugate the incorporated modifications to a number of substrates to carry out SDS-PAGE, fluorescence microscopy, and enrichment for LC-MS/MS. Using LC-MS/MS, we identified 103 and 174 proteins from the myristic and palmitic azide enrichments, with 27 and 45 proteins respectively that differentiated HIV-1 infected from uninfected cells. This approach has provided us with important insights into HIV-1 biology and is widely applicable to many virological systems.

  17. Changing the Culture of Academic Medicine: Critical Mass or Critical Actors?

    Science.gov (United States)

    Helitzer, Deborah L; Newbill, Sharon L; Cardinali, Gina; Morahan, Page S; Chang, Shine; Magrane, Diane

    2017-05-01

    By 2006, women constituted 34% of academic medical faculty, reaching a critical mass. Theoretically, with critical mass, culture and policy supportive of gender equity should be evident. We explore whether having a critical mass of women transforms institutional culture and organizational change. Career development program participants were interviewed to elucidate their experiences in academic health centers (AHCs). Focus group discussions were held with institutional leaders to explore their perceptions about contemporary challenges related to gender and leadership. Content analysis of both data sources revealed points of convergence. Findings were interpreted using the theory of critical mass. Two nested domains emerged: the individual domain included the rewards and personal satisfaction of meaningful work, personal agency, tensions between cultural expectations of family and academic roles, and women's efforts to work for gender equity. The institutional domain depicted the sociocultural environment of AHCs that shaped women's experience, both personally and professionally, lack of institutional strategies to engage women in organizational initiatives, and the influence of one leader on women's ascent to leadership. The predominant evidence from this research demonstrates that the institutional barriers and sociocultural environment continue to be formidable obstacles confronting women, stalling the transformational effects expected from achieving a critical mass of women faculty. We conclude that the promise of critical mass as a turning point for women should be abandoned in favor of "critical actor" leaders, both women and men, who individually and collectively have the commitment and power to create gender-equitable cultures in AHCs.

  18. Critically damped quantum search.

    Science.gov (United States)

    Mizel, Ari

    2009-04-17

    Although measurement and unitary processes can accomplish any quantum evolution in principle, thinking in terms of dissipation and damping can be powerful. We propose a modification of Grover's algorithm in which the idea of damping plays a natural role. Remarkably, we find that there is a critical damping value that divides between the quantum O(sqrt[N]) and classical O(N) search regimes. In addition, by allowing the damping to vary in a fashion we describe, one obtains a fixed-point quantum search algorithm in which ignorance of the number of targets increases the number of oracle queries only by a factor of 1.5.

  19. Critically damped quantum search

    OpenAIRE

    Mizel, Ari

    2008-01-01

    Although measurement and unitary processes can accomplish any quantum evolution in principle, thinking in terms of dissipation and damping can be powerful. We propose a modification of Grover's algorithm in which the idea of damping plays a natural role. Remarkably, we have found that there is a critical damping value that divides between the quantum $O(\\sqrt{N})$ and classical O(N) search regimes. In addition, by allowing the damping to vary in a fashion we describe, one obtains a fixed-poin...

  20. Critical cyberculture studies

    CERN Document Server

    Silver, David; Jones, Steve

    2006-01-01

    Starting in the early 1990s, journalists and scholars began responding to and trying to take account of new technologies and their impact on our lives. By the end of the decade, the full-fledged study of cyberculture had arrived. Today, there exists a large body of critical work on the subject, with cutting-edge studies probing beyond the mere existence of virtual communities and online identities to examine the social, cultural, and economic relationships that take place online. Taking stock of the exciting work that is being done and positing what cyberculture's future might look like, Criti

  1. Critical Points of Contact

    DEFF Research Database (Denmark)

    Jensen, Ole B.; Morelli, Nicola

    2011-01-01

    where the networks meet and establish contact. Thus we argue for the usefulness of the notion of Critical Point of Contact (CPC) to deepen our understanding of the actual life within networks. En route to this notion we draw upon theories within as diverse realms such as interaction design, service...... design, geography, and mobility studies. After the introduction in section we develop and define the notion of CPC based upon a broad set of disciplines and theories. We illustrate the usefulness of the notion within the field of mobility in the network city and within the field of service design...

  2. Critical Zen art history

    Directory of Open Access Journals (Sweden)

    Gregory P. A. Levin

    2016-12-01

    Full Text Available This essay sketches a history of the study of Zen art from the late nineteenth century to post-war reconsiderations, leading towards what I term “critical Zen art studies.” The latter, I suggest, has been undertaken by historians of art and others to challenge normative definitions of Zen art based on modern constructs, revise understanding of the types and functions of visual art important to Chan/Sŏn/Zen Buddhist monasteries, and study iconographies and forms not as a transparent aesthetic indices to Zen Mind or No Mind but as rhetorically, ritually, and socially complex, even unruly, events of representation.

  3. Durability of critical infrastructures

    Directory of Open Access Journals (Sweden)

    Raluca Pascu

    2011-08-01

    Full Text Available The paper deals with those infrastructures by which world society, under the pressure ofdemographic explosion, self-survives. The main threatening comes not from terrorist attacks, but fromthe great natural catastrophes and global climate change. It’s not for the first time in history when suchmeasures of self-protection are built up. First objective of this paper is to present the background fordurability analysis. Then, with the aid of these mathematical tools the absolute durability of three linearmodels, typical for critical infrastructures, are successively calculated. In order to enhance the durabilityof critical infrastructures the solution based on redundancies is chosen. Five types of connection theredundancies for each of the three models are considered. Three topological schemes for connecting theredundancies are adopted: locally, by twining and globally. Absolute values of durability in all fifteenmodels with redundancies are further calculated. Then, the relative performances of enhanced durabilityin the same fifteen models, compared with the three original models, considered as references, areanalysed. The relative costs of the same fifteen models and in similar topologic conditions are furtheranalysed. By dividing the performance with cost the relative profitableness of each model is obtained.Finally, the three initial models, each reshaped with redundancies in three selective modes, arecompared from the perspective of their relative profitableness. The outcomes of this paper are original.They are of practical interests in planning the maintenance programs and checking the plausibility ofproposed interventions according to the clause 7.4 of ISO 13822:2001

  4. Design of Critical Components

    Science.gov (United States)

    Hendricks, Robert C.; Zaretsky, Erwin V.

    2001-01-01

    Critical component design is based on minimizing product failures that results in loss of life. Potential catastrophic failures are reduced to secondary failures where components removed for cause or operating time in the system. Issues of liability and cost of component removal become of paramount importance. Deterministic design with factors of safety and probabilistic design address but lack the essential characteristics for the design of critical components. In deterministic design and fabrication there are heuristic rules and safety factors developed over time for large sets of structural/material components. These factors did not come without cost. Many designs failed and many rules (codes) have standing committees to oversee their proper usage and enforcement. In probabilistic design, not only are failures a given, the failures are calculated; an element of risk is assumed based on empirical failure data for large classes of component operations. Failure of a class of components can be predicted, yet one can not predict when a specific component will fail. The analogy is to the life insurance industry where very careful statistics are book-kept on classes of individuals. For a specific class, life span can be predicted within statistical limits, yet life-span of a specific element of that class can not be predicted.

  5. Review - Critical Han Studies

    Directory of Open Access Journals (Sweden)

    Zhiguo Ye

    2013-12-01

    Full Text Available Review of: Thomas Mullaney, James Leibold, Stéphane Gros, and Eric Armand Vanden Bussche (eds. 2012. Critical Han Studies: The History, Representation, and Identity of China's Majority. Berkeley, Calif: University of California Press. This path-breaking volume is an academic collaboration that emerged out of the "Critical Han Studies Conference and Workshop" at Stanford University in April 2008. Eleven scholars contributed to the question of what it means to be 'Han' in China, both historically and at present. Constituting over ninety percent of China's population, the Han are not only the largest ethnic group in China, but are also one of the largest categories of collective identity in the world. Despite this, the dominant Han group has so far eluded careful scholarly scrutiny, with the Han often referred to as an unmarked majority category in contemporary China. This volume challenges such conventional views by conceptualizing new interdisciplinary approaches to the question of Hanness. The eleven essays of the volume are divided into three themes: 'Han and China', 'The Problem of Han Origins', and 'The Problem of Han Formations'. The first theme, comprised of four essays, analyzes the ties that bind the category of Han to those of Chinese ethnicity, race, and polity. Kevin Carrico in "Recentering China: The Cantonese in and Beyond the Han" questions a single, unitary Hanness that he believes conceals "countless other perceived and imagined lines of differentiation" (25. The study examines how multiple identities...

  6. Writer, Reader, Critic: Comparing Critical Theories as Discourse.

    Science.gov (United States)

    de Beaugrande, Robert

    1984-01-01

    Attempts to show how each of three influential critical theories--deconstructionism, reader response criticism, and authorial intention--implies a particular view of how literary discourse is or should be processed and indicates that each view is in part justified, but not to the extent claimed by the critics themselves. (CRH)

  7. What Makes Critical Thinking Critical for Adult ESL Students

    Science.gov (United States)

    Miekley, Joshua P.

    2014-01-01

    Critical-thinking skills help to prepare adult education students for a successful transition to college degree programs and for job advancement. Yet fostering critical thinking poses a challenge to ESL instructors. Brookfield (2012) provides a way forward for adult educators when he explains that the crux of critical thinking is to discover one's…

  8. Critical Literature Pedagogy: Teaching Canonical Literature for Critical Literacy

    Science.gov (United States)

    Borsheim-Black, Carlin; Macaluso, Michael; Petrone, Robert

    2014-01-01

    This article introduces Critical Literature Pedagogy (CLP), a pedagogical framework for applying goals of critical literacy within the context of teaching canonical literature. Critical literacies encompass skills and dispositions to understand, question, and critique ideological messages of texts; because canonical literature is often…

  9. Conserved nonlocal dynamics and critical behavior of uranium ferromagnetic superconductors.

    Science.gov (United States)

    Singh, Rohit; Dutta, Kishore; Nandy, Malay K

    2017-01-01

    A recent theoretical study [Phys. Rev. Lett. 112, 037202 (2014)10.1103/PhysRevLett.112.037202] has revealed that systems such as uranium ferromagnetic superconductors obey conserved dynamics. To capture the critical behavior near the paramagnetic to ferromagnetic phase transition of these compounds, we study the conserved critical dynamics of a nonlocal Ginzburg-Landau model. A dynamic renormalization-group calculation at one-loop order yields the critical indices in the leading order of ε=d_{c}-d, where d_{c}=4-2ρ is the upper critical dimension, with ρ an exponent in the nonlocal interaction. The predicted static critical exponents are found to be comparable with the available experimentally observed critical exponents for strongly uniaxial uranium ferromagnetic superconductors. The corresponding dynamic exponent z and linewidth exponent w are found to be z=4-ρε/4+O(ε^{2}) and w=1+ρ+3ε/4+O(ε^{2}).

  10. Conserved nonlocal dynamics and critical behavior of uranium ferromagnetic superconductors

    Science.gov (United States)

    Singh, Rohit; Dutta, Kishore; Nandy, Malay K.

    2017-01-01

    A recent theoretical study [Phys. Rev. Lett. 112, 037202 (2014), 10.1103/PhysRevLett.112.037202] has revealed that systems such as uranium ferromagnetic superconductors obey conserved dynamics. To capture the critical behavior near the paramagnetic to ferromagnetic phase transition of these compounds, we study the conserved critical dynamics of a nonlocal Ginzburg-Landau model. A dynamic renormalization-group calculation at one-loop order yields the critical indices in the leading order of ɛ =dc-d , where dc=4 -2 ρ is the upper critical dimension, with ρ an exponent in the nonlocal interaction. The predicted static critical exponents are found to be comparable with the available experimentally observed critical exponents for strongly uniaxial uranium ferromagnetic superconductors. The corresponding dynamic exponent z and linewidth exponent w are found to be z =4 -ρ ɛ /4 +O (ɛ2) and w =1 +ρ +3 ɛ /4 +O (ɛ2) .

  11. Puerto Rico Revealed Preference data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Revealed preference models provide insights into recreational angler behavior and the economic value of recreational fishing trips. Revealed preference data is...

  12. Endangered Species Act Critical Habitat

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Critical habitat (CH) is designated for the survival and recovery of species listed as threatened or endangered under the Endangered Species Act (ESA). Critical...

  13. Modeling Dependencies in Critical Infrastructures

    NARCIS (Netherlands)

    Nieuwenhuijs, A.H.; Luiijf, H.A.M.; Klaver, M.H.A.

    2009-01-01

    This paper describes a model for expressing critical infrastructure dependencies. The model addresses the limitations of existing approaches with respect to clarity of definition, support for quality and the influence of operating states of critical infrastructures and environmental factors.

  14. Mission Critical Occupation (MCO) Charts

    Data.gov (United States)

    Office of Personnel Management — Agencies report resource data and targets for government-wide mission critical occupations and agency specific mission critical and/or high risk occupations. These...

  15. Revealing the Beast Within

    Science.gov (United States)

    2003-07-01

    Deeply Embedded Massive Stellar Clusters Discovered in Milky Way Powerhouse Summary Peering into a giant molecular cloud in the Milky Way galaxy - known as W49 - astronomers from the European Southern Observatory (ESO) have discovered a whole new population of very massive newborn stars . This research is being presented today at the International Astronomical Union's 25th General Assembly held in Sydney, Australia, by ESO-scientist João Alves. With the help of infrared images obtained during a period of excellent observing conditions with the ESO 3.5-m New Technology Telescope (NTT) at the La Silla Observatory (Chile), the astronomers looked deep into this molecular cloud and discovered four massive stellar clusters, with hot and energetic stars as massive as 120 solar masses. The exceedingly strong radiation from the stars in the largest of these clusters is "powering" a 20 light-year diameter region of mostly ionized hydrogen gas (a "giant HII region"). W49 is one of the most energetic regions of star formation in the Milky Way. With the present discovery, the true sources of the enormous energy have now been revealed for the first time, finally bringing to an end some decades of astronomical speculations and hypotheses. PR Photo 21a/03 : Colour Composite of W49A (NTT+SOFI). PR Photo 21b/03 : Radio and Near-Infrared Composite of W49A Giant molecular clouds Stars form predominantly inside Giant Molecular Clouds which populate our Galaxy, the Milky Way. One of the most prominent of these is W49 , which has a mass of a million solar masses. It is located some 37,000 light-years away and is the most luminous star-forming region known in our home galaxy: its luminosity is several million times the luminosity of our Sun. A smaller region within this cloud is denoted W49A - this is one of the strongest radio-emitting areas known in the Galaxy . Massive stars are excessive in all ways. Compared to their smaller and ligther brethren, they form at an Olympic speed and

  16. Systems analysis of MVA-C induced immune response reveals its significance as a vaccine candidate against HIV/AIDS of clade C.

    Directory of Open Access Journals (Sweden)

    Carmen Elena Gómez

    Full Text Available Based on the partial efficacy of the HIV/AIDS Thai trial (RV144 with a canarypox vector prime and protein boost, attenuated poxvirus recombinants expressing HIV-1 antigens are increasingly sought as vaccine candidates against HIV/AIDS. Here we describe using systems analysis the biological and immunological characteristics of the attenuated vaccinia virus Ankara strain expressing the HIV-1 antigens Env/Gag-Pol-Nef of HIV-1 of clade C (referred as MVA-C. MVA-C infection of human monocyte derived dendritic cells (moDCs induced the expression of HIV-1 antigens at high levels from 2 to 8 hpi and triggered moDCs maturation as revealed by enhanced expression of HLA-DR, CD86, CD40, HLA-A2, and CD80 molecules. Infection ex vivo of purified mDC and pDC with MVA-C induced the expression of immunoregulatory pathways associated with antiviral responses, antigen presentation, T cell and B cell responses. Similarly, human whole blood or primary macrophages infected with MVA-C express high levels of proinflammatory cytokines and chemokines involved with T cell activation. The vector MVA-C has the ability to cross-present antigens to HIV-specific CD8 T cells in vitro and to increase CD8 T cell proliferation in a dose-dependent manner. The immunogenic profiling in mice after DNA-C prime/MVA-C boost combination revealed activation of HIV-1-specific CD4 and CD8 T cell memory responses that are polyfunctional and with effector memory phenotype. Env-specific IgG binding antibodies were also produced in animals receiving DNA-C prime/MVA-C boost. Our systems analysis of profiling immune response to MVA-C infection highlights the potential benefit of MVA-C as vaccine candidate against HIV/AIDS for clade C, the prevalent subtype virus in the most affected areas of the world.

  17. What Is Critical Discourse Analysis?

    Directory of Open Access Journals (Sweden)

    Gavin Kendall

    2007-05-01

    Full Text Available In this interview, Ruth WODAK discusses the beginnings of her career, and what propelled her into critical discourse analysis. She analyses what makes critical discourse analysis "critical", distinguishes criticalness from dogmatism, but expounds upon the relationship between critique and norms. Finally, she discusses how "integrative indisciplinarity" might help us with problems of disciplinary incommensurability. URN: urn:nbn:de:0114-fqs0702297

  18. Critical Incidents in Internal Relationships

    OpenAIRE

    Voima, Päivi

    2000-01-01

    Critical incidents have had an important role in service quality and service management research. The focus of critical-incident studies has gradually shifted from separate acts and episodes towards relationships, and even switching from one relationship to another. The Critical Incident Technique has mainly been used when studying the service sector, concentrating on the customer's perception of critical incidents. Although some studies have considered the perceptions of employees important,...

  19. Teaching Critical Thinking

    CERN Document Server

    Holmes, N G; Bonn, D A

    2015-01-01

    The ability to make decisions based on data, with its inherent uncertainties and variability, is a complex and vital skill in the modern world. The need for such quantitative critical thinking occurs in many different contexts, and while it is an important goal of education, that goal is seldom being achieved. We argue that the key element for developing this ability is repeated practice in making decisions based on data, with feedback on those decisions. We demonstrate a structure for providing suitable practice that can be applied in any instructional setting that involves the acquisition of data and relating that data to scientific models. This study reports the results of applying that structure in an introductory physics lab course. Students in an experimental condition were repeatedly instructed to make and act on quantitative comparisons between datasets, and between data and models, an approach that is common to all science disciplines. These instructions were slowly faded across the course. After the...

  20. [Nutrition in critical illness].

    Science.gov (United States)

    Ökrös, Ilona

    2014-12-21

    Critically ill patients are often unable to eat by themselves over a long period of time, sometimes for weeks. In the acute phase, serious protein-energy malnutrition may develop with progressive muscle weakness, which may result in assisted respiration of longer duration as well as longer stay in intensive care unit and hospital. In view of the metabolic processes, energy and protein intake targets should be defined and the performance of metabolism should be monitored. Enteral nutrition is primarily recommended. However, parenteral supplementation is often necessary because of the disrupted tolerance levels of the gastrointestinal system. Apparently, an early parenteral supplementation started within a week would be of no benefit. Some experts believe that muscle loss can be reduced by increased target levels of protein. Further studies are needed on the effect of immune system feeding, fatty acids and micronutrients.

  1. Critical anthropometry for menarche.

    Science.gov (United States)

    Gonzales, G F; Villena, A

    1996-08-01

    The objective of the study was to determine if any anthropometric measurements, including weight, height, and upper arm circumference, or the calculated body mass index observed at the time of menarche may be used as a threshold for menarche. The sample was randomly selected from 1133 girls aged 10-18 years from two groups, one residing in Lima, Peru, at 150 m above sea level, and the second in Cerro de Pasco, Peru, at 4340 m above sea level. For the purpose of this study, all girls who reached menarche at least 1 month before the study (n = 93) were considered as cases (girls at menarche); 88 girls without menarche at the time of the study were randomly matched by age and place of residence, and designated as controls. All subjects came from the same low socioeconomic status. Body weight at menarche was 44.6 +/- 5.09 kg (mean +/- SD; coefficient of variation [CV], 11.4%). Height at menarche was 151.6 +/- 5.5 cm (CV, 3.7%). Body mass index, defined as weight/height2.15, was 18.26 +/- 1.96 kg/m2.15 at menarche (CV, 10.7%). Upper arm circumference at menarche was 21.8 +/- 1.6 cm (CV, 7.3%). The logistic regression analysis showed that any of the four anthropometric measurements analyzed were critical for menarche; i.e., any of the four could be used as a threshold for menarche. Data from the present study do not support the hypothesis that there are anthropometric markers that are critical for menarche.

  2. Why Critical design is useless

    DEFF Research Database (Denmark)

    Ebbesen, Toke Riis

    practical use, critical design may then become another echo chamber for designers, where they can safely repeat the slogans of design modernism without changing the world. The aim of this presentation will be to return to critical design in the light of the concept of use, and thus to critically examine...

  3. From Memorization to Critical Thinking

    Institute of Scientific and Technical Information of China (English)

    Long; Feng

    2015-01-01

    The essay introduces the author’s experiences in the UK and Canada to explain the difference of educational methods in China and western countries:from memorization to critical thinking.The author explains what the critical thinking is and what features should a critical thinker have,then give some suggestions to improve Chinese educational system.

  4. Critical Quantitative Inquiry in Context

    Science.gov (United States)

    Stage, Frances K.; Wells, Ryan S.

    2014-01-01

    This chapter briefly traces the development of the concept of critical quantitative inquiry, provides an expanded conceptualization of the tasks of critical quantitative research, offers theoretical explanation and justification for critical research using quantitative methods, and previews the work of quantitative criticalists presented in this…

  5. Critical Quantitative Inquiry in Context

    Science.gov (United States)

    Stage, Frances K.; Wells, Ryan S.

    2014-01-01

    This chapter briefly traces the development of the concept of critical quantitative inquiry, provides an expanded conceptualization of the tasks of critical quantitative research, offers theoretical explanation and justification for critical research using quantitative methods, and previews the work of quantitative criticalists presented in this…

  6. What Is Critical about Sociology?

    Science.gov (United States)

    Buechler, Steven

    2008-01-01

    Critical thinking is often presented as a generic technique. This article develops an alternative that links critique more closely to the sociological perspective. I suggest three answers to the above question: that the sociological perspective is critical for comprehending complex issues, that all sociology is implicitly critical by virtue of its…

  7. A Critically Reflective Social Studies?

    Science.gov (United States)

    Clarke, Marge

    1990-01-01

    Examines social studies in the twenty-first century from a critical theory perspective. Traces critical reflection's origins from Marxist educational theories to Jurgen Habermas's critical theory. Highlights Fred Newmann's curricular model, "Education for Citizen Action," for developing competent action in public affairs. Advocates infusing…

  8. A Critically Reflective Social Studies?

    Science.gov (United States)

    Clarke, Marge

    1990-01-01

    Examines social studies in the twenty-first century from a critical theory perspective. Traces critical reflection's origins from Marxist educational theories to Jurgen Habermas's critical theory. Highlights Fred Newmann's curricular model, "Education for Citizen Action," for developing competent action in public affairs. Advocates…

  9. Focus groups reveal consumer ambivalence.

    Science.gov (United States)

    1983-01-01

    According to qualitative research, Salvadoreans are ambivalent about the use of contraceptives. Since complete responsibility for management of the CSM project was accepted by the Association Demografica Salvadorena (ADS), the agency which operates the contraceptive social marketing project in El Salvador, in November 1980, the need for decisions in such areas as product price increases, introduction of new condom brands, promotion of the vaginal foaming tablet, and assessment of product sales performance had arisen. The ICSMP funded market research, completed during 1983, was intended to provide the data on which such decisions by ADS could be based. The qualitative research involved 8 focus groups, comprised of men and women, aged 18-45, contraceptive users and nonusers, from the middle and lower socioeconomic strata of the city of San Salvador and other suburban areas. In each group a moderator led discussion of family planning and probed respondents for specific attitudes, knowledge, and behavior regarding the use of contraceptives. To assess attitudes at a more emotional level, moderators asked respondents to "draw" their ideas on certain issues. A marked discrepancy was revealed between respondents' intellectual responses to the issues raised in group discussion, as opposed to their feelings expressed in the drawings. Intellectually, participants responded very positively to family planning practice, but when they were asked to draw their perceptions, ambivalent feelings emerged. Drawings of both the user and the nonuser convey primarily negative aspects for either choice. The user is tense and moody toward her children; the nonuser loses her attractiveness and "dies." Figures also show drawings of some of the attitudes of single and married male participants. 1 drawing shows an incomplete and a complete circle, symbolizing a sterilized man (incomplete) and a nonsterilized man (complete). Another picture depicts a chained man who has lost his freedom

  10. Myopathies in critical illness: characterization and nutritional aspects.

    Science.gov (United States)

    Burnham, Ellen L; Moss, Marc; Ziegler, Thomas R

    2005-07-01

    Myopathies related to critical illness have received increasing recognition over the past decade and are common in patients even after a brief period in the intensive care unit. Recent studies have revealed that myopathies in the critically ill may in fact be more prevalent than neuropathies and that morbidity and mortality may be greater. Protein catabolism, an increase in urinary nitrogen loss, and muscle wasting are observed in critical illness myopathy. Muscle biopsies in critically ill patients demonstrate low glutamine levels, low protein/DNA levels, and high concentrations of extracellular water. The increased flux of glutamine in muscle in these patients is thought to be insufficient to meet the body's requirement for glutamine, and thus in critical illness this amino acid may be classified as "conditionally essential." Three subtypes of critical illness myopathy have been described that are often grouped together as acute quadriplegic myopathy: thick-filament myopathy, critical illness myopathy, and necrotizing myopathy. These can be differentiated based on clinical features and muscle biopsy. Treatments for critical illness myopathies range from primary prevention, i.e., avoiding myopathy-inducing drugs, to novel nutritional therapies, such as glutamine and glutathione supplementation. One should be particularly vigilant for the development of myopathies in critically ill alcoholic patients, who may have a chronic alcoholic myopathy at baseline. In the past decade, advances have been made in characterizing and identifying patients with myopathies due to critical illness. However, additional studies must be performed in order to develop appropriate therapies for this potentially devastating disorder.

  11. Applying critical thinking to nursing.

    Science.gov (United States)

    Price, Bob

    2015-08-19

    Critical thinking and writing are skills that are not easy to acquire. The term 'critical' is used differently in social and clinical contexts. Nursing students need time to master the inquisitive and ruminative aspects of critical thinking that are required in academic environments. This article outlines what is meant by critical thinking in academic settings, in relation to both theory and reflective practice. It explains how the focus of a question affects the sort of critical thinking required and offers two taxonomies of learning, to which students can refer when analysing essay requirements. The article concludes with examples of analytical writing in reference to theory and reflective practice.

  12. Towards critical aspects of Confucianism

    Directory of Open Access Journals (Sweden)

    Dunaj Ľubomír

    2016-12-01

    Full Text Available This paper consists of two parts. The first deals with the issue of whether it is possible to coherently employ the term ‘critical Confucian’ in general, i.e. whether it is a paradox or oxymoron. It will be argued that Confucianism should not be identified with any particular ideology and, therefore, can be critical. This critical potential, in turn, can be developed by bringing it into dialogue with Critical Theory. As such, the second part indicates, in an introductory way, some possible overlaps between Confucianism and Critical Theory by comparing Heiner Roetz’s and Axel Honneth’s respective interpretations of these traditions.

  13. Enhancing superconducting critical current by randomness.

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Y. L.; Thoutam, L. R.; Xiao, Z. L.; Shen, B.; Pearson, J.; Divan, R.; Ocola, L. E.; Crabtree, G. W.; Kwok, W. K.

    2016-01-11

    The key ingredient of high critical currents in a type-II superconductor is defect sites that pin vortices. Here, we demonstrate that a random pinscape, an overlooked pinning system in nanopatterned superconductors, can lead to a substantially larger critical current enhancement at high magnetic fields than an ordered array of vortex pin sites. We reveal that the better performance of a random pinscape is due to the variation of the local density of its pinning sites, which mitigates the motion of vortices. This is confirmed by achieving even higher enhancement of the critical current through a conformally mapped random pinscape, where the distribution of the local density of pinning sites is further enlarged. Our findings highlight the potential of random pinscapes in enhancing the superconducting critical currents of applied superconductors in which random pin sites of nanoscale defects emerging in the materials synthesis process or through ex-situ irradiation are the only practical choice for large-scale production. Our results may also stimulate research on effects of a random pinscape in other complementary systems such as colloidal crystals, Bose-Einstein condensates, and Luttinger liquids.

  14. Are Earthquakes a Critical Phenomenon?

    Science.gov (United States)

    Ramos, O.

    2014-12-01

    Earthquakes, granular avalanches, superconducting vortices, solar flares, and even stock markets are known to evolve through power-law distributed events. During decades, the formalism of equilibrium phase transition has coined these phenomena as critical, which implies that they are also unpredictable. This work revises these ideas and uses earthquakes as the paradigm to demonstrate that slowly driven systems evolving through uncorrelated and power-law distributed avalanches (UPLA) are not necessarily critical systems, and therefore not necessarily unpredictable. By linking the correlation length to the pdf of the distribution, and comparing it with the one obtained at a critical point, a condition of criticality is introduced. Simulations in the classical Olami-Feder-Christensen (OFC) earthquake model confirm the findings, showing that earthquakes are not a critical phenomenon. However, one single catastrophic earthquake may show critical properties and, paradoxically, the emergence of this temporal critical behaviour may eventually carry precursory signs of catastrophic events.

  15. A Holographic Model For Quantum Critical Responses

    CERN Document Server

    Myers, Robert C; Witczak-Krempa, William

    2016-01-01

    We analyze the dynamical response functions of strongly interacting quantum critical states described by conformal field theories (CFTs). We construct a self-consistent holographic model that incorporates the relevant scalar operator driving the quantum critical phase transition. Focusing on the finite temperature dynamical conductivity $\\sigma(\\omega,T)$, we study its dependence on our model parameters, notably the scaling dimension of the relevant operator. It is found that the conductivity is well-approximated by a simple ansatz proposed by Katz et al [1] for a wide range of parameters. We further dissect the conductivity at large frequencies $\\omega >> T$ using the operator product expansion, and show how it reveals the spectrum of our model CFT. Our results provide a physically-constrained framework to study the analytic continuation of quantum Monte Carlo data, as we illustrate using the O(2) Wilson-Fisher CFT. Finally, we comment on the variation of the conductivity as we tune away from the quantum cri...

  16. Gadamer’s Critical Views of Husserl

    Directory of Open Access Journals (Sweden)

    Alireza

    2015-10-01

    Full Text Available usserl, besides Dilthey and Heidegger, is one of the most important philosophers who had big influence on Gadamer and is the philosopher who is frequently referred to in his works. Gadamer’s encounter with Husserl, in spite of his greatness in the eyes of Gadamer, was always a hermeneutical one. Considering and criticizing Husselr’s phenomenology, Gadamer tries to discover his unspoken thoughts. Few scholars have even considered Gadamer and Husserl’s relationships, which among them much more are people who tried to trace the influence of Husserl in Gadamer’s philosophical hermeneutics. Instead, here, we will reveal and highlight the Gadamer’s critical views of the key concepts of Husserl's phenomenology such as subjectivity, pure perception, intersubjectivity, transcendental ego, and alter ego. We believe Gadamer practically remained loyal to his hermeneutical approach. Instead of just repeating the thoughts, he goes behind them and opens new horizons.

  17. Critical heat flux thermodynamics

    Energy Technology Data Exchange (ETDEWEB)

    Collado, F.J. E-mail: fjk@posta.unizar.es

    2002-11-01

    Convective boiling in subcooled water flowing through a heated channel is essential in many engineering applications where high heat flux need to be accommodated, such as in the divertor plates of fusion reactors. There are many available correlations for predicting heat transfer in the individual regimes of the empirical Nukiyama boiling curve, although unfortunately there is no physical fundamentals of such curve. Recently, the author has shown that the classical entropy balance could contain key information about boiling heat transfer. So, it was found that the average thermal gap in the heated channel (the wall temperature minus the average temperature of the coolant fluid) was strongly correlated with the efficiency of a theoretical reversible engine placed in this thermal gap. In this work and from the new proposed correlation, a new expression of the wall temperature in function of the average fluid temperature is derived and successfully checked against experimental data from General Electric. This expression suggests a new and simple definition of the critical heat flux (CHF), a key parameter of the thermal-hydraulic design of fusion reactors. Finally, based on the new definition, the CHF trends are commented.

  18. Horizon as Critical Phenomenon

    CERN Document Server

    Lee, Sung-Sik

    2016-01-01

    We show that renormalization group(RG) flow can be viewed as a gradual wave function collapse, where a quantum state associated with the action of field theory evolves toward a final state that describes an IR fixed point. The process of collapse is described by the radial evolution in the dual holographic theory. If the theory is in the same phase as the assumed IR fixed point, the initial state is smoothly projected to the final state. If in a different phase, the initial state undergoes a phase transition which in turn gives rise to a horizon in the bulk geometry. We demonstrate the connection between critical behavior and horizon in an example, by deriving the bulk metrics that emerge in various phases of the U(N) vector model in the large N limit based on the holographic dual constructed from quantum RG. The gapped phase exhibits a geometry that smoothly ends at a finite proper distance in the radial direction. The geometric distance in the radial direction measures a complexity : the depth of RG transfo...

  19. Chiropractic: a critical evaluation.

    Science.gov (United States)

    Ernst, Edzard

    2008-05-01

    Chiropractic was defined by D.D. Palmer as "a science of healing without drugs." About 60,000 chiropractors currently practice in North America, and, worldwide, billions are spent each year for their services. This article attempts to critically evaluate chiropractic. The specific topics include the history of chiropractic; the internal conflicts within the profession; the concepts of chiropractic, particularly those of subluxation and spinal manipulation; chiropractic practice and research; and the efficacy, safety, and cost of chiropractic. A narrative review of selected articles from the published chiropractic literature was performed. For the assessment of efficacy, safety, and cost, the evaluation relied on previously published systematic reviews. Chiropractic is rooted in mystical concepts. This led to an internal conflict within the chiropractic profession, which continues today. Currently, there are two types of chiropractors: those religiously adhering to the gospel of its founding fathers and those open to change. The core concepts of chiropractic, subluxation and spinal manipulation, are not based on sound science. Back and neck pain are the domains of chiropractic but many chiropractors treat conditions other than musculoskeletal problems. With the possible exception of back pain, chiropractic spinal manipulation has not been shown to be effective for any medical condition. Manipulation is associated with frequent mild adverse effects and with serious complications of unknown incidence. Its cost-effectiveness has not been demonstrated beyond reasonable doubt. The concepts of chiropractic are not based on solid science and its therapeutic value has not been demonstrated beyond reasonable doubt.

  20. Collimation of fast electrons in critical density plasma channel

    OpenAIRE

    2015-01-01

    Significantly collimated fast electron beam with a divergence angle 10° (FWHM) is generated through the interaction of ultra-intense laser light with a uniform critical density plasma in experiments and 2D PIC simulations. In the experiment, the uniform critical density plasma is created by ionizing an ultra-low density foam target. The spacial distribution of the fast electron is observed by Imaging Plate. 2D PIC simulation and post process analysis reveal magnetic collimation of energetic e...

  1. “Systematic Approach New Criticism under criticism: focusing on

    OpenAIRE

    پیروز پیروز

    2009-01-01

    New Criticism focuses on rigorous analysis of poetry and literature, irrespective of peripheral issues. Its fundamental tenet is the autonomy of literary texts. Therefore, New Criticism shuns dealing with such peripheral issues as authors’ biography, social, political and historical events; since this approach considers all these issues as non-artistic. In principle, the followers of New Criticism subscribe to closed analysis; because, only semantic and aesthetic dimensions are important for ...

  2. 75 FR 5146 - Hewlett Packard Company Business Critical Systems, Mission Critical Business Software Division...

    Science.gov (United States)

    2010-02-01

    ... Employment and Training Administration Hewlett Packard Company Business Critical Systems, Mission Critical... Business Critical Systems, Mission Critical Business Software Division, OpenVMS Operating System... of Hewlett Packard Company, Business Critical Systems, Mission Critical Business Software Division...

  3. 75 FR 11918 - Hewlett Pachard Company, Business Critical Systems, Mission Critical Business Software Division...

    Science.gov (United States)

    2010-03-12

    ... Employment and Training Administration Hewlett Pachard Company, Business Critical Systems, Mission Critical... Company, Business Critical Systems, Mission Critical Business Software Division, Openvms Operating System..., applicable to workers of Hewlett Packard Company, Business Critical Systems, Mission Critical Business...

  4. A Critical Discourse Analysis of News Discourse

    Institute of Scientific and Technical Information of China (English)

    王冬梅

    2008-01-01

    This paper aims to explore the underlying ideologies as well as the relationship between news discoutse and ideology by analyzing a news report on Saddam Hussei's capture in New York Times from the perspective of critical discourse analysis.While making the analysis,this paper studies the details of linguistic features,trausitivity,modality and themstic choice which are based on the three metafuoctions of language proposed by Halliday in systemic-functional grammar, so as to reveal how news discourse is used by powerful groups to reinforce dominant ideologies.and to gain profound insights about the stance and attitude maintained in news reports.

  5. Neuronal plasticity: beyond the critical period.

    Science.gov (United States)

    Hübener, Mark; Bonhoeffer, Tobias

    2014-11-06

    Neuronal plasticity in the brain is greatly enhanced during critical periods early in life and was long thought to be rather limited thereafter. Studies in primary sensory areas of the neocortex have revealed a substantial degree of plasticity in the mature brain, too. Often, plasticity in the adult neocortex lies dormant but can be reactivated by modifications of sensory input or sensory-motor interactions, which alter the level and pattern of activity in cortical circuits. Such interventions, potentially in combination with drugs targeting molecular brakes on plasticity present in the adult brain, might help recovery of function in the injured or diseased brain.

  6. Critical Discourse Analysis of“Leftover Ladies”

    Institute of Scientific and Technical Information of China (English)

    余媛

    2013-01-01

    In recent years, many new words have emerged in our life, such as “leftover men and ladies.”The investigation and reports of the media and the TV series have attracted a great deal of attention. Taking a critical discourse analysis perspective(CDA), this article analyzes textual data from some news reports and interviews related to this heated topic. Norman Fairclough’s Discourse and Social Change would provide some theoretical framework for this study. The ifndings reveal how this concept is constructed and how it inlfuences our society.

  7. Critical incident analysis through narrative reflective practice: A case study

    Directory of Open Access Journals (Sweden)

    Thomas S. C. Farrell

    2013-01-01

    Full Text Available Teachers can reflect on their practices by articulating and exploring incidents they consider critical to themselves or others. By talking about these critical incidents, teachers can make better sense of seemingly random experiences that occur in their teaching because they hold the real inside knowledge, especially personal intuitive knowledge, expertise and experience that is based on their accumulated years as language educators teaching in schools and classrooms. This paper is about one such critical incident analysis that an ESL teacher in Canada revealed to her critical friend and how both used McCabe’s (2002 narrative framework for analyzing an important critical incident that occurred in the teacher’s class.

  8. Critical energy deficit and mortality in critically ill patients

    Directory of Open Access Journals (Sweden)

    Marcia Carolina Siqueira-Paese

    Full Text Available Objective: We investigate the influence of caloric and protein deficit on mortality and length of hospital stay of critically ill patients. Methods: A cohort prospective study including 100 consecutive patients in a tertiary intensive care unit (ICU receiving enteral or parenteral nutrition. The daily caloric and protein deficit were collected each day for a maximum of 30 days. Energy deficits were divided into critical caloric deficit (≥ 480 kcal/day and non-critical caloric deficit (≤ 480 kcal/day; and in critical protein deficit (≥ 20 g/day and non-critical protein deficit (≤ 20 g/day. The findings were correlated with hospital stay and mortality. Results: The mortality rate was 33%. Overall, the patients received 65.4% and 67.7% of the caloric and protein needs. Critical caloric deficit was found in 72% of cases and critical protein deficit in 70% of them. There was a significant correlation between length of stay and accumulated caloric deficit (R = 0.37; p < 0.001 and protein deficit (R = 0.28; p < 0.001. The survival analysis showed that mortality was greater in patients with both critical caloric (p < 0.001 and critical protein deficits (p < 0.01. The Cox regression analysis showed that critical protein deficit was associated with higher mortality (HR 0.25, 95% CI 0.07-0.93, p = 0.03. Conclusions: The incidence of caloric and protein deficit in the ICU is high. Both caloric and protein deficits increase the length of hospital stay, and protein deficit greater than 20 g/day is an independent factor for mortality in critical care unit.

  9. Post-Industrial Cultural Criticism

    DEFF Research Database (Denmark)

    Kammer, Aske

    2015-01-01

    Integrating perspectives from research into cultural and post-industrial journalism, this article presents a pilot study of websites with reviews of arts and culture conducted by amateurs. Such websites constitute a popular space for cultural criticism, and one that challenges traditional...... reviewers have highly specialized knowledge of culture and, on that basis, argues that the emergence of this type of critic might represent a qualitative strengthening of cultural criticism....

  10. Evaluation of Saxton critical experiments

    Energy Technology Data Exchange (ETDEWEB)

    Joo, Hyung Kook; Noh, Jae Man; Jung, Hyung Guk; Kim, Young Il; Kim, Young Jin [Korea Atomic Energy Research Institute, Taejon (Korea, Republic of)

    1997-12-31

    As a part of International Criticality Safety Benchmark Evaluation Project (ICSBEP), SAXTON critical experiments were reevaluated. The effects of k{sub eff} of the uncertainties in experiment parameters, fuel rod characterization, soluble boron, critical water level, core structure, {sup 241}Am and {sup 241}Pu isotope number densities, random pitch error, duplicated experiment, axial fuel position, model simplification, etc., were evaluated and added in benchmark-model k{sub eff}. In addition to detailed model, the simplified model for Saxton critical experiments was constructed by omitting the top, middle, and bottom grids and ignoring the fuel above water. 6 refs., 1 fig., 3 tabs. (Author)

  11. Protein requirement in critical illness.

    Science.gov (United States)

    Hoffer, Leonard John

    2016-05-01

    How much protein do critically ill patients require? For the many decades that nutritional support has been used there was a broad consensus that critically ill patients need much more protein than required for normal health. Now, however, some clinical investigators recommend limiting all macronutrient provision during the early phase of critical illness. How did these conflicting recommendations emerge? Which of them is correct? This review explains the longstanding recommendation for generous protein provision in critical illness, analyzes the clinical trials now being claimed to refute it, and concludes with suggestions for clinical investigation and practice.

  12. An introduction to critical paths.

    Science.gov (United States)

    Coffey, Richard J; Richards, Janet S; Remmert, Carl S; LeRoy, Sarah S; Schoville, Rhonda R; Baldwin, Phyllis J

    2005-01-01

    A critical path defines the optimal sequencing and timing of interventions by physicians, nurses, and other staff for a particular diagnosis or procedure. Critical paths are developed through collaborative efforts of physicians, nurses, pharmacists, and others to improve the quality and value of patient care. They are designed to minimize delays and resource utilization and to maximize quality of care. Critical paths have been shown to reduce variation in the care provided, facilitate expected outcomes, reduce delays, reduce length of stay, and improve cost-effectiveness. The approach and goals of critical paths are consistent with those of total quality management (TQM) and can be an important part of an organization's TQM process.

  13. Quantum criticality from Fisher information

    Science.gov (United States)

    Song, Hongting; Luo, Shunlong; Fu, Shuangshuang

    2017-04-01

    Quantum phase transition is primarily characterized by a qualitative sudden change in the ground state of a quantum system when an external or internal parameter of the Hamiltonian is continuously varied. Investigating quantum criticality using information-theoretic methods has generated fruitful results. Quantum correlations and fidelity have been exploited to characterize the quantum critical phenomena. In this work, we employ quantum Fisher information to study quantum criticality. The singular or extremal point of the quantum Fisher information is adopted as the estimated thermal critical point. By a significant model constructed in Quan et al. (Phys Rev Lett 96: 140604, 2006), the effectiveness of this method is illustrated explicitly.

  14. Universal Off-Equilibrium Scaling of Critical Cumulants in the QCD Phase Diagram

    Science.gov (United States)

    Mukherjee, Swagato; Venugopalan, Raju; Yin, Yi

    2016-11-01

    Exploiting the universality between the QCD critical point and the three-dimensional Ising model, closed form expressions derived for nonequilibrium critical cumulants on the crossover side of the critical point reveal that they can differ in both magnitude and sign from equilibrium expectations. We demonstrate here that key elements of the Kibble-Zurek framework of nonequilibrium phase transitions can be employed to describe the dynamics of these critical cumulants. Our results suggest that observables sensitive to critical dynamics in heavy-ion collisions should be expressible as universal scaling functions, thereby providing powerful model-independent guidance in searches for the QCD critical point.

  15. Do Critical Thinking Exercises Improve Critical Thinking Skills?

    Science.gov (United States)

    Cotter, Ellen M.; Tally, Carrie Sacco

    2009-01-01

    Although textbooks routinely include exercises to improve critical thinking skills, the effectiveness of these exercises has not been closely examined. Additionally, the connection between critical thinking skills and formal operational thought is also relatively understudied. In the study reported here, college students completed measures of…

  16. The Critical Advantage: Developing Critical Thinking Skills in School

    Science.gov (United States)

    Gormley, William T., Jr.

    2017-01-01

    In "The Critical Advantage," noted scholar and early childhood expert William T. Gormley, Jr. takes a wide ranging look at the important role of critical thinking in preparing students for college, careers, and civic life. Drawing on research from psychology, philosophy, business, political science, neuroscience, and other disciplines,…

  17. Critical Language Awareness. Part II: Towards Critical Alternatives.

    Science.gov (United States)

    Clark, Romy; And Others

    1991-01-01

    The notion of critical linguistics is examined, and it is argued that the diverse objectives usually given for Language Awareness programs appear to be given desocializing weightings in actual materials. Ways that Critical Language Awareness can be incorporated into a family history writing project are described. (20 references) (Author/LB)

  18. Marxian Critical Theory and Educational Administration: A Criticism.

    Science.gov (United States)

    Willower, Donald J.

    The critical examination of society's circumstances, institutions, and cultural predispositions and values can help detect problems and injustices and can suggest opportunities for improvement. Neo-Marxist critical theorists of the Frankfurt school are committed to work of this kind, but their perspective is informed by Marxian precepts, and their…

  19. Neutron Imaging Reveals Internal Plant Hydraulic Dynamics

    Energy Technology Data Exchange (ETDEWEB)

    Warren, Jeffrey [ORNL; Bilheux, Hassina Z [ORNL; Kang, Misun [ORNL; Voisin, Sophie [ORNL; Cheng, Chu-Lin [ORNL; Horita, Jusuke [ORNL; Perfect, Edmund [ORNL

    2013-01-01

    Many terrestrial ecosystem processes are constrained by water availability and transport within the soil. Knowledge of plant water fluxes is thus critical for assessing mechanistic processes linked to biogeochemical cycles, yet resolution of root structure and xylem water transport dynamics has been a particularly daunting task for the ecologist. Through neutron imaging, we demonstrate the ability to non-invasively monitor individual root functionality and water fluxes within Zea mays L. (maize) and Panicum virgatum L. (switchgrass) seedlings growing in a sandy medium. Root structure and growth were readily imaged by neutron radiography and neutron computed tomography. Seedlings were irrigated with water or deuterium oxide and imaged through time as a growth lamp was cycled on to alter leaf demand for water. Sub-millimeter scale resolution reveals timing and magnitudes of root water uptake, redistribution within the roots, and root-shoot hydraulic linkages, relationships not well characterized by other techniques.

  20. Chemotaxis: new role for Ras revealed

    Institute of Scientific and Technical Information of China (English)

    Jianshe Yan; Dale Hereld; Tian Jin

    2010-01-01

    @@ A recent study of chemotaxis revealed a new role for the proto-oncogene Ras in the social ameba Dictyostelium discoideum.Chemotaxis,the directional movement of cells toward chemokines and other chemoattractants,plays critical roles in diverse physiological processes,such as mobilization of immune cells to fight invading microorganisms,targeting of metastatic cancer cells to specific tissues,and guidance of sperm cells to ova during fertilization.This work,published in the July 26 issue of The Journal of Cell Biology,was conducted in Dr.Devreotes' lab at John Hopkins University and Dr.Parent's lab at National Cancer Institute.This research team demonstrated that RasC functions as an upstream regulator of TORC2 and thereby governs the effects of TORC2-PKB signaling on the cytoskeleton and cell migration.

  1. Thinking Critically about Critical Thinking: Integrating Online Tools to Promote Critical Thinking

    OpenAIRE

    B. Jean Mandernach

    2006-01-01

    The value and importance of critical thinking is clearly established; the challenge for instructors lies in successfully promoting students’ critical thinking skills within the confines of a traditional classroom experience. Since instructors are faced with limited student contact time to meet their instructional objectives and facilitate learning, they are often forced to make instructional decisions between content coverage, depth of understanding, and critical analysis of course materia...

  2. Critical market crashes

    Science.gov (United States)

    Sornette, D.

    2003-04-01

    patterns have been documented for essentially all crashes on developed as well as emergent stock markets, on currency markets, on company stocks, and so on. We review this discovery at length and demonstrate how to use this insight and the detailed predictions obtained from these models to forecast crashes. For this, we review the major crashes of the past that occurred on the major stock markets of the planet and describe the empirical evidence of the universal nature of the critical log-periodic precursory signature of crashes. The concept of an “anti-bubble” is also summarized, with the Japanese collapse from the beginning of 1991 to present, taken as a prominent example. A prediction issued and advertised in January 1999 has been until recently born out with remarkable precision, predicting correctly several changes of trends, a feat notoriously difficult using standard techniques of economic forecasting. We also summarize a very recent analysis the behavior of the U.S. S&P500 index from 1996 to August 2002 and the forecast for the two following years. We conclude by presenting our view of the organization of financial markets.

  3. Critical Thinking: Schemata vs. Skills.

    Science.gov (United States)

    Brandhorst, Allan R.

    1989-01-01

    Refutes the idea that critical thinking is not a skill by analyzing it from the phenomenological perspective of Edmund Husserl, and from the hermeneutic perspective of Martin Heidegger. Develops the thesis that critical thinking is a restructuring of schemata. Addresses the problem of attention or student engagement. (LS)

  4. Critical Pedagogy in Health Education

    Science.gov (United States)

    Matthews, Catherine

    2014-01-01

    Objective: This review investigated how the three-phase model of critical pedagogy, based on the writings of Paulo Freire, can be put into practice in health education. Design: The study considers literature related to the fields of health education, health promotion and critical pedagogy. Setting: The study is a scholarly review completed as part…

  5. Construction of YBS Critical Assembly

    Institute of Scientific and Technical Information of China (English)

    ZHOU; Qi; LIANG; Shu-hong; ZHU; Qing-fu; ZHANG; Wei; YANG; Li-jun; QUAN; Yan-hui

    2015-01-01

    Supported by the XDA program of CAS,the YBS critical assembly has been constructed for the experimental research of the coupling and influence characteristics of spallation target and reactor in ADS system.This work consists of two parts:one is the conversion of the reactor hall and control room,and the other manufacture,installation and commissioning of the critical

  6. Critical Thinking: Schemata vs. Skills.

    Science.gov (United States)

    Brandhorst, Allan R.

    1989-01-01

    Refutes the idea that critical thinking is not a skill by analyzing it from the phenomenological perspective of Edmund Husserl, and from the hermeneutic perspective of Martin Heidegger. Develops the thesis that critical thinking is a restructuring of schemata. Addresses the problem of attention or student engagement. (LS)

  7. Quantifying Learning in Critical Thinking

    Science.gov (United States)

    Fliegel, Richard; Holland, John

    2013-01-01

    This article describes a three-year study assessing change in critical thinking demonstrated in essays written for regular class assignments. A rubric was designed and scorers trained to assess critical thinking holistically without knowledge of the writing prompt or author's status. The longitudinal improvement in scores earned by freshmen over…

  8. Scrutiny of Critical Thinking Concept

    Science.gov (United States)

    Atabaki, Ali Mohammad Siahi; Keshtiaray, Narges; Yarmohammadian, Mohammad H.

    2015-01-01

    Learning critical thinking skills are the goal of educational systems so the term "critical thinking" (CT) is frequently found in educational policy documents. Despite this frequency, however, precise understandings among teachers of what CT really means do not exit. The present study is designed to answer the following question. We can…

  9. Teaching Students to Think Critically

    Science.gov (United States)

    Black, Susan

    2005-01-01

    In this article, the author stresses that teachers need to teach their students to think critically and to reason their way. One prerequisite for teaching critical thinking is a classroom climate of high expectations, teacher warmth and encouragement, and pleasant physical surroundings. Schools should see to it that students become progressively…

  10. Harnessing Critical Incidents for Learning

    Science.gov (United States)

    Patahuddin, Sitti Maesuri; Lowrie, Tom

    2015-01-01

    A critical incident is a situation or event that holds significance for learning, both for the students and teachers. This paper presents four examples of critical incidents from a Year 7 teacher's lesson excerpts in Indonesia involving teaching of fractions, to show how they shaped classroom situation, brought forward elements of conflict, and…

  11. Critical Thinking: Implications for Research.

    Science.gov (United States)

    Craver, Kathleen W.

    1989-01-01

    Synthesizes critical thinking research in four areas--reading, writing, group interactions, and speaking--and discusses the implications of those studies for library information science research. The potential for employing critical thinking strategies in bibliographic instruction and the need for library science educators to initiate their own…

  12. Quantifying Learning in Critical Thinking

    Science.gov (United States)

    Fliegel, Richard; Holland, John

    2013-01-01

    This article describes a three-year study assessing change in critical thinking demonstrated in essays written for regular class assignments. A rubric was designed and scorers trained to assess critical thinking holistically without knowledge of the writing prompt or author's status. The longitudinal improvement in scores earned by freshmen over…

  13. Course Design for Critical Thinking.

    Science.gov (United States)

    Furedy, John J.; Furedy, Christine

    1979-01-01

    A fourth year honors thesis research course in psychology at the University of Toronto uses the device of adversarial interaction to improve critical thinking. Course components, including thesis submission, research seminar, student relations, and supervision, are designed to simulate the constraints, criticism, and relationships of actual…

  14. The Importance of Critical Literacy

    Science.gov (United States)

    Janks, Hilary

    2012-01-01

    This paper is divided into three parts. It begins by making an argument for the ongoing importance of critical literacy at a moment when there are mutterings about its being passe. The second part of the paper formulates the argument with the use of illustrative texts. It concludes with examples of critical literacy activities that I argue, are…

  15. Traditional Literacy and Critical Thinking

    Science.gov (United States)

    Dando, Priscille

    2016-01-01

    How school librarians focus on activating critical thinking through traditional literacy development can proactively set the stage for the deep thinking that occurs in all literacy development. The critical-thinking skills students build while becoming accomplished readers and writers provide the foundation for learning in a variety of…

  16. The Metaethics of Critical Theories

    NARCIS (Netherlands)

    Stahl, Titus; Thompson, Michael

    2017-01-01

    Critical theories, from their beginning in Marx’s philosophy to the Frankfurt School with its different generations, have always been characterized by a certain ambivalence toward moral questions. They often conceive themselves as an alternative to traditional moral philosophy, which is criticized

  17. Critical factors for EIA implementation

    DEFF Research Database (Denmark)

    Zhang, Jasmine; Kørnøv, Lone; Christensen, Per

    2013-01-01

    between different critical factors and how they relate to different stages in the EIA and thus influence the decision making process. After reviewing 33 refereed journal articles published between 1999 and 2011, we identified 203 notions of critical factors. Of these, 102 related to different stages...

  18. Critical Thinking in Physical Education

    Science.gov (United States)

    Humphries, Charlotte

    2014-01-01

    Changes in American education require that teachers are evaluated more often, and expectations increasingly include teaching to develop critical thinking skills. This article uses Bloom's taxonomy in describing ways physical educators can include critical thinking in their lessons, both to enhance their teaching and to meet expectations of…

  19. Critical thinking in physics education

    Science.gov (United States)

    Sadidi, Farahnaz

    2016-07-01

    We agree that training the next generation of leaders of the society, who have the ability to think critically and form a better judgment is an important goal. It is a long-standing concern of Educators and a long-term desire of teachers to establish a method in order to teach to think critically. To this end, many questions arise on three central aspects: the definition, the evaluation and the design of the course: What is Critical Thinking? How can we define Critical Thinking? How can we evaluate Critical Thinking? Therefore, we want to implement Critical Thinking in physics education. How can we teach for Critical Thinking in physics? What should the course syllabus and materials be? We present examples from classical physics and give perspectives for astro-particle physics. The main aim of this paper is to answer the questions and provide teachers with the opportunity to change their classroom to an active one, in which students are encouraged to ask questions and learn to reach a good judgment. Key words: Critical Thinking, evaluation, judgment, design of the course.

  20. Applying critical analysis - main methods

    Directory of Open Access Journals (Sweden)

    Miguel Araujo Alonso

    2012-02-01

    Full Text Available What is the usefulness of critical appraisal of literature? Critical analysis is a fundamental condition for the correct interpretation of any study that is subject to review. In epidemiology, in order to learn how to read a publication, we must be able to analyze it critically. Critical analysis allows us to check whether a study fulfills certain previously established methodological inclusion and exclusion criteria. This is frequently used in conducting systematic reviews although eligibility criteria are generally limited to the study design. Critical analysis of literature and be done implicitly while reading an article, as in reading for personal interest, or can be conducted in a structured manner, using explicit and previously established criteria. The latter is done when formally reviewing a topic.

  1. Critical Thinking in Higher Education: Unfulfilled Expectations

    Directory of Open Access Journals (Sweden)

    Mehrdad Rezaee

    2012-05-01

    Full Text Available Success in adult life and effective functioning in education depends among other things on critical thinking. The present study consisted of two parts. First, critical thinking (CT skill of a group of 68 students majoring in education in Islamic Azad University, Kermanshah Branch was evaluated. The participants, divided into two experimental and control groups, received California Critical Thinking Skills Test (CCTST which is a 34 item Multiple-Choice test. The students in the control group were freshmen and the experimental group, junior students. To the researchers’ dismay, junior education students did not perform significantly better than did the freshman students. Using a qualitative method of research, another study was conducted to see whether the university instructors in the education department who had the responsibility of teaching different courses to the same students were aware of the principles of CT. A semi-structured interview was conducted and eight volunteering faculty members in the department of education took part in the interview. Result revealed that, although these instructors highly valued CT and were aware of its tenets, there were some constraints which did provide a situation to let the students practice CT in their classrooms, and much had to be done to help instructors implement CT in their classrooms.

  2. Investigating Adolescents' Critical Consciousness Development through a Character Framework.

    Science.gov (United States)

    Seider, Scott; Tamerat, Jalene; Clark, Shelby; Soutter, Madora

    2017-02-16

    Brazilian philosopher-educator Paulo Freire defined critical consciousness as the ability to engage in reflection and action upon the world in order to transform it. A growing body of research has found that critical consciousness is predictive of a number of important academic and civic outcomes in adolescents from oppressed groups. The present mixed methods study considered the critical consciousness development of 335 adolescents (57% female, 92% African American or Latinx) attending urban secondary schools that sought to foster their students' critical consciousness, but featured five different pedagogical approaches. We hypothesized that considering these adolescents' critical consciousness development through a character lens would highlight ways in which different schooling models contribute differentially to their students' development of the intellectual, performance, and civic dimensions of critical consciousness. Longitudinal analyses revealed significant differences in the critical consciousness development of adolescents attending different schooling models along these dimensions. Interviews with adolescents and field work conducted at their schools offered insight into the programming and practices that may have contributed to these differences in students' critical consciousness development.

  3. Critical crack path assessments in failure investigations

    Directory of Open Access Journals (Sweden)

    Robert D. Caligiuri

    2015-10-01

    Full Text Available This paper presents a case study in which identification of the controlling crack path was critical to identifying the root cause of the failure. The case involves the rupture of a 30-inch (0.76 m natural gas pipeline in 2010 that tragically led to the destruction of a number of homes and the loss of life. The segment of the pipeline that ruptured was installed in 1956. The longitudinal seam of the segment that ruptured was supposed to have been fabricated by double submerged arc welding. Unfortunately, portions of the segment only received a single submerged arc weld on the outside, leaving unwelded areas on the inside diameter. Post-failure examination of the segment revealed that the rupture originated at one of these unwelded areas. Examination also revealed three additional crack paths or zones emanating from the unwelded area: a zone of ductile tearing, a zone of fatigue, and a zone of cleavage fracture, in that sequence. Initial investigators ignored the ductile tear, assumed the critical crack path was the fatigue component, and (incorrectly concluded that the root cause of the incident was the failure of the operator to hydrotest the segment after it was installed in 1956. However, as discussed in this paper, the critical path or mechanism was the ductile tear. Furthermore, it was determined that the ductile tear was created during the hydrotest at installation by a mechanism known as pressure reversal. Thus the correct root cause of the rupture was the hydrotest the operator subjected the segment to at installation, helping to increase the awareness of operators and regulators about the potential problems associated with hydrotesting.

  4. Genetic characterization of ebi reveals its critical role in Drosophila wing growth

    Science.gov (United States)

    Walker, Cherryl D; Orme, Mariam H; Leevers, Sally J

    2011-01-01

    The ebi gene of Drosophila melanogaster has been implicated in diverse signaling pathways, cellular functions and developmental processes. However, a thorough genetic analysis of this gene has been lacking and the true extent of its biological roles is unclear. Here, we characterize eleven ebi mutations and find that ebi has a novel role in promoting growth of the wing imaginal disc: viable combinations of mutant alleles give rise to adults with small wings. Wing discs with reduced EBI levels are correspondingly small and exhibit downregulation of Notch target genes. Furthermore, we show that EBI colocalizes on polytene chromosomes with Smrter (SMR), a transcriptional corepressor, and Suppressor of Hairless (SU(H)), the primary transcription factor involved in Notch signaling. Interestingly, the mammalian orthologs of ebi, transducin β-like 1 (TBL1) and TBL-related 1 (TBLR1), function as corepressor/coactivator exchange factors and are required for transcriptional activation of Notch target genes. We hypothesize that EBI acts to activate (de-repress) transcription of Notch target genes important for Drosophila wing growth by functioning as a corepressor/coactivator exchange factor for SU(H). PMID:22041576

  5. Transcriptional dynamics reveal critical roles for non-coding RNAs in the immediate-early response.

    Directory of Open Access Journals (Sweden)

    Stuart Aitken

    2015-04-01

    Full Text Available The immediate-early response mediates cell fate in response to a variety of extracellular stimuli and is dysregulated in many cancers. However, the specificity of the response across stimuli and cell types, and the roles of non-coding RNAs are not well understood. Using a large collection of densely-sampled time series expression data we have examined the induction of the immediate-early response in unparalleled detail, across cell types and stimuli. We exploit cap analysis of gene expression (CAGE time series datasets to directly measure promoter activities over time. Using a novel analysis method for time series data we identify transcripts with expression patterns that closely resemble the dynamics of known immediate-early genes (IEGs and this enables a comprehensive comparative study of these genes and their chromatin state. Surprisingly, these data suggest that the earliest transcriptional responses often involve promoters generating non-coding RNAs, many of which are produced in advance of canonical protein-coding IEGs. IEGs are known to be capable of induction without de novo protein synthesis. Consistent with this, we find that the response of both protein-coding and non-coding RNA IEGs can be explained by their transcriptionally poised, permissive chromatin state prior to stimulation. We also explore the function of non-coding RNAs in the attenuation of the immediate early response in a small RNA sequencing dataset matched to the CAGE data: We identify a novel set of microRNAs responsible for the attenuation of the IEG response in an estrogen receptor positive cancer cell line. Our computational statistical method is well suited to meta-analyses as there is no requirement for transcripts to pass thresholds for significant differential expression between time points, and it is agnostic to the number of time points per dataset.

  6. Meta-analysis reveals a critical period for management of powdery mildew on Hop cones

    Science.gov (United States)

    Results of 28 field trials conducted over a 12-year period investigating management of hop powdery mildew caused by Podosphaera macularis were quantitatively summarized by meta-analysis to compare product efficacy and use patterns by mode of action as defined by Fungicide Resistance Action Committe...

  7. Comparative genomic analysis reveals a critical role of de novo nucleotide biosynthesis for Saccharomyces cerevisiae virulence.

    Directory of Open Access Journals (Sweden)

    Roberto Pérez-Torrado

    Full Text Available In recent years, the number of human infection cases produced by the food related species Saccharomyces cerevisiae has increased. Whereas many strains of this species are considered safe, other 'opportunistic' strains show a high degree of potential virulence attributes and can cause infections in immunocompromised patients. Here we studied the genetic characteristics of selected opportunistic strains isolated from dietary supplements and also from patients by array comparative genomic hybridization. Our results show increased copy numbers of IMD genes in opportunistic strains, which are implicated in the de novo biosynthesis of the purine nucleotides pathway. The importance of this pathway for virulence of S. cerevisiae was confirmed by infections in immunodeficient murine models using a GUA1 mutant, a key gene of this pathway. We show that exogenous guanine, an end product of this pathway in its triphosphorylated form, increases the survival of yeast strains in ex vivo blood infections. Finally, we show the importance of the DNA damage response that activates dNTP biosynthesis in yeast cells during ex vivo blood infections. We conclude that opportunistic yeasts may use an enhanced de novo biosynthesis of the purine nucleotides pathway to increase survival and favor infections in the host.

  8. Domain complementation studies reveal residues critical for the activity of the mannitol permease from Escherichia coli.

    Science.gov (United States)

    Vos, Erwin P P; ter Horst, Ramon; Poolman, Bert; Broos, Jaap

    2009-02-01

    This paper presents domain complementation studies in the mannitol transporter, EIImtl, from Escherichia coli. EIImtl is responsible for the transport and concomitant phosphorylation of mannitol over the cytoplasmic membrane. By using tryptophan-less EIImtl as a basis, each of the four phenylalanines located in the cytoplasmic loop between putative transmembrane helices II and III in the membrane-embedded C domain were replaced by tryptophan, yielding the mutants W97, W114, W126, and W133. Except for W97, these single-tryptophan mutants exhibited a high, wild-type-like, binding affinity for mannitol. Of the four mutants, only W114 showed a high mannitol phosphorylation activity. EIImtl is functional as a dimer and the effect of these mutations on the oligomeric activity was investigated via heterodimer formation (C/C domain complementation studies). The low phosphorylation activities of W126 and W133 could be increased 7-28 fold by forming heterodimers with either the C domain of W97 (IICmtlW97) or the inactive EIImtl mutant G196D. W126 and W133, on the other hand, did not complement each other. This study points towards a role of positions 97, 126 and 133 in the oligomeric activation of EIImtl. The involvement of specific residue positions in the oligomeric functioning of a sugar-translocating EII protein has not been presented before.

  9. Genomic Characterization of Esophageal Squamous Cell Carcinoma Reveals Critical Genes Underlying Tumorigenesis and Poor Prognosis

    Science.gov (United States)

    Qin, Hai-De; Liao, Xiao-Yu; Chen, Yuan-Bin; Huang, Shao-Yi; Xue, Wen-Qiong; Li, Fang-Fang; Ge, Xiao-Song; Liu, De-Qing; Cai, Qiuyin; Long, Jirong; Li, Xi-Zhao; Hu, Ye-Zhu; Zhang, Shao-Dan; Zhang, Lan-Jun; Lehrman, Benjamin; Scott, Alan F.; Lin, Dongxin; Zeng, Yi-Xin; Shugart, Yin Yao; Jia, Wei-Hua

    2016-01-01

    The genetic mechanisms underlying the poor prognosis of esophageal squamous cell carcinoma (ESCC) are not well understood. Here, we report somatic mutations found in ESCC from sequencing 10 whole-genome and 57 whole-exome matched tumor-normal sample pairs. Among the identified genes, we characterized mutations in VANGL1 and showed that they accelerated cell growth in vitro. We also found that five other genes, including three coding genes (SHANK2, MYBL2, FADD) and two non-coding genes (miR-4707-5p, PCAT1), were involved in somatic copy-number alterations (SCNAs) or structural variants (SVs). A survival analysis based on the expression profiles of 321 individuals with ESCC indicated that these genes were significantly associated with poorer survival. Subsequently, we performed functional studies, which showed that miR-4707-5p and MYBL2 promoted proliferation and metastasis. Together, our results shed light on somatic mutations and genomic events that contribute to ESCC tumorigenesis and prognosis and might suggest therapeutic targets. PMID:27058444

  10. Reaction-diffusion-like formalism for plastic neural networks reveals dissipative solitons at criticality

    Science.gov (United States)

    Grytskyy, Dmytro; Diesmann, Markus; Helias, Moritz

    2016-06-01

    Self-organized structures in networks with spike-timing dependent synaptic plasticity (STDP) are likely to play a central role for information processing in the brain. In the present study we derive a reaction-diffusion-like formalism for plastic feed-forward networks of nonlinear rate-based model neurons with a correlation sensitive learning rule inspired by and being qualitatively similar to STDP. After obtaining equations that describe the change of the spatial shape of the signal from layer to layer, we derive a criterion for the nonlinearity necessary to obtain stable dynamics for arbitrary input. We classify the possible scenarios of signal evolution and find that close to the transition to the unstable regime metastable solutions appear. The form of these dissipative solitons is determined analytically and the evolution and interaction of several such coexistent objects is investigated.

  11. Internal Domains of Natural Porous Media Revealed: Critical Locations for Transport, Storage, and Chemical Reaction

    Energy Technology Data Exchange (ETDEWEB)

    Zachara, John M.; Brantley, Susan L.; Chorover, Jon D.; Ewing, Robert P.; Kerisit, Sebastien N.; Liu, Chongxuan; Perfect, E.; Rother, Gernot; Stack, Andrew G.

    2016-03-16

    Internal pore domains exist within rocks, lithic fragments, subsurface sediments and soil aggregates. These domains, which we term internal domains in porous media (IDPM), contain a significant fraction of their porosity as nanopores, dominate the reactive surface area of diverse porous media types, and are important locations for chemical reactivity and hydrocarbon storage. Traditionally difficult to interrogate, advances in instrumentation and imaging methods are providing new insights on the physical structures and chemical attributes of IDPM. In this review we: discuss analytical methods to characterize IDPM, evaluate what has been learned about their size distributions, connectivity, and extended structures; determine whether they exhibit unique chemical reactivity; and assess potential for their inclusion in reactive transport models. Three key findings are noteworthy. 1) A combination of methods now allows complete characterization of the porosity spectrum of natural materials and its connectivity; while imaging microscopies are providing three dimensional representations of the interconnected pore network. 2) Chemical reactivity in pores <10 nm is expected to be different from micro and macropores, yet research performed to date is inconclusive on the nature, direction, and magnitude of effect. 3) Existing continuum reactive transport models treat IDPM as a sub-grid feature with average, empirical, scale-dependent parameters; and are not formulated to include detailed information on pore networks. Overall we find that IDPM are key features controlling hydrocarbon release from shales in hydrofracking systems, organic matter stabilization and recalcitrance in soil, weathering and soil formation, and long term inorganic and organic contaminant behavior in the vadose zone and groundwater. We conclude with an assessment of impactful research opportunities to advance understanding of IDPM, and to incorporate their important effects in reactive transport models for improved environmental simulation and prediction.

  12. Reaction-diffusion-like formalism for plastic neural networks reveals dissipative solitons at criticality.

    Science.gov (United States)

    Grytskyy, Dmytro; Diesmann, Markus; Helias, Moritz

    2016-06-01

    Self-organized structures in networks with spike-timing dependent synaptic plasticity (STDP) are likely to play a central role for information processing in the brain. In the present study we derive a reaction-diffusion-like formalism for plastic feed-forward networks of nonlinear rate-based model neurons with a correlation sensitive learning rule inspired by and being qualitatively similar to STDP. After obtaining equations that describe the change of the spatial shape of the signal from layer to layer, we derive a criterion for the nonlinearity necessary to obtain stable dynamics for arbitrary input. We classify the possible scenarios of signal evolution and find that close to the transition to the unstable regime metastable solutions appear. The form of these dissipative solitons is determined analytically and the evolution and interaction of several such coexistent objects is investigated.

  13. New inducible genetic method reveals critical roles of GABA in the control of feeding and metabolism

    Science.gov (United States)

    Currently available inducibleCre/loxPsystems, despite their considerable utility in gene manipulation, have pitfalls in certain scenarios, such as unsatisfactory recombination rates and deleterious effects on physiology and behavior. To overcome these limitations, we designed a new, inducible gene-t...

  14. “Systematic Approach New Criticism under criticism: focusing on

    Directory of Open Access Journals (Sweden)

    پیروز پیروز

    2009-01-01

    Full Text Available New Criticism focuses on rigorous analysis of poetry and literature, irrespective of peripheral issues. Its fundamental tenet is the autonomy of literary texts. Therefore, New Criticism shuns dealing with such peripheral issues as authors’ biography, social, political and historical events; since this approach considers all these issues as non-artistic. In principle, the followers of New Criticism subscribe to closed analysis; because, only semantic and aesthetic dimensions are important for them and texts are seen more like closed and insulated systems. The writer of this article introduces systematic theories and their characteristics. Literary texts are seen as a system, based on the systematic models. The writer has also attempted to examine the ‘analytic and mechanistic’ dimensions of New Criticism, as well as the interaction of sub-systems with meta-systems in literary texts.

  15. Critical Pedagogy and Democratic Education: Possibilities for Cross-Pollination

    Science.gov (United States)

    Edwards, D. Brent, Jr.

    2010-01-01

    Reviewing the literature on critical pedagogy (CP) and democratic education (DE) reveals that very little has been written comparing the two (Knight and Pearl in Urban Rev 32(2):197-226, 2000). After reading the Urban Review article by Knight and Pearl (2000)--the only publication explicitly comparing the two approaches to education--I was…

  16. LITERARY CRITICISM AND THE TEACHING OF THE NOVEL.

    Science.gov (United States)

    LUNDY, JACK T.

    CONTEMPORARY NOVELS CAN BE USED EFFECTIVELY BY TEACHERS IN THE CLASSROOM. JOHN KNOWLES'"A SEPARATE PEACE," FOR EXAMPLE, LENDS ITSELF TO DIFFERENT KINDS OF LITERARY CRITICISM. AN ARCHETYPAL INTERPRETATION OF KNOWLES' THEMATIC PURPOSE REVEALS THAT THE NOVEL IS BASED ON TWO MYTHS--THE INITIATION OF INNOCENCE INTO EXPERIENCE AND THE…

  17. Critical Pedagogy and Democratic Education: Possibilities for Cross-Pollination

    Science.gov (United States)

    Edwards, D. Brent, Jr.

    2010-01-01

    Reviewing the literature on critical pedagogy (CP) and democratic education (DE) reveals that very little has been written comparing the two (Knight and Pearl in Urban Rev 32(2):197-226, 2000). After reading the Urban Review article by Knight and Pearl (2000)--the only publication explicitly comparing the two approaches to education--I was…

  18. Using Learning Styles in Information Literacy: Critical Considerations for Librarians

    Science.gov (United States)

    Sanderson, Heather

    2011-01-01

    Librarians are using learning styles as a tool to engage students and enhance their teaching. However, a review of the literature reveals that learning styles theory is complex and problematic. It is important to base our practice on sound pedagogy. This critical examination of learning styles explores the issues surrounding them and what they can…

  19. A Critical Perspective of Contemporary Unemployment Policy and Practices

    Science.gov (United States)

    Blustein, David L.; Medvide, Mary Beth; Wan, Carol M.

    2012-01-01

    This article explores the challenges of unemployment via the lens of critical psychology. The conventional discourse on unemployment is critiqued, revealing ways in which conventional policies and practices serve to further marginalize the lives of the unemployed and impede the development of ethical, effective, and empathic individual…

  20. The Role of Critical Formalism in Music Education

    Science.gov (United States)

    Louth, J. Paul

    2012-01-01

    This article discusses the emancipatory potential of critical formalism, a mode of critique that may be helpful in revealing to music students the taken-for-granted nature of some common musical and educational notions whose socially constructed nature may not always appear evident. The work is presented in two parts: "theory" and "praxis." The…

  1. Sleep disturbances in critically ill patients in ICU

    DEFF Research Database (Denmark)

    Boyko, Yuliya; Ording, H; Jennum, P

    2012-01-01

    Sleep disturbances in the intensive care unit (ICU) seem to lead to development of delirium, prolonged ICU stay, and increased mortality. That is why sufficient sleep is important for good outcome and recovery in critically ill patients. A variety of small studies reveal pathological sleep patterns...

  2. Using Learning Styles in Information Literacy: Critical Considerations for Librarians

    Science.gov (United States)

    Sanderson, Heather

    2011-01-01

    Librarians are using learning styles as a tool to engage students and enhance their teaching. However, a review of the literature reveals that learning styles theory is complex and problematic. It is important to base our practice on sound pedagogy. This critical examination of learning styles explores the issues surrounding them and what they can…

  3. Toward a possible next geomagnetic critical transition?

    Science.gov (United States)

    Qamili, Enkelejda; De Santis, Angelo; Wu, Lixin

    2014-05-01

    The geomagnetic field is subject to possible reversals or excursions of polarity during its temporal evolution. Considering the characteristics of the recent geomagnetic field, a possible imminent geomagnetic reversal or excursion would not be completely unexpected. In that case, such a phenomenon would represent one of the very few natural hazards that are really global. The South Atlantic Anomaly (SAA) is a great depression of the geomagnetic field strength at the Earth's surface, caused by a reverse magnetic flux in the terrestrial outer core. In analogy with critical point phenomena characterized by some cumulative quantity, we fit the surface extent of this anomaly over the last 400 yr with power law or logarithmic functions in reverse time, also decorated by log-periodic oscillations, whose final singularity (a critical point tc) reveals a great change in the near future (2034±3 yr), when the SAA area reaches almost a hemisphere. An interesting aspect that has recently been found is the possible direct connection between the SAA and the global mean sea level (GSL). That the GSL is somehow connected with SAA is also confirmed by the similar result when an analogous critical-like fit is performed over GSL: the corresponding critical point (2033±11 yr) agrees, within the estimated errors, with the value found for the SAA. From this result, we point out the intriguing conjecture that tc would be the time of no return, after which the geomagnetic field could fall into an irreversible process of a global geomagnetic transition that could be a reversal or excursion of polarity.

  4. Being Critical in Marketing Studies

    DEFF Research Database (Denmark)

    Dholakia, Nikilesh

    2012-01-01

    In this article, I argue that an elevated macro-level perspective is imperative for conducting critical studies in the fields of marketing and consumer research. There are epistemic barriers to operating in this manner, and I offer several suggestions for overcoming these barriers. Finally, I...... review the research spaces for critical studies of marketing in various global settings and conclude that United Kingdom and Nordic Europe have the best epistemic climate, and this region needs to take leadership in promoting greater range of macro and critical studies of marketing in the rest...

  5. Being Critical in Marketing Studies

    DEFF Research Database (Denmark)

    Dholakia, Nikilesh

    2012-01-01

    review the research spaces for critical studies of marketing in various global settings and conclude that United Kingdom and Nordic Europe have the best epistemic climate, and this region needs to take leadership in promoting greater range of macro and critical studies of marketing in the rest......In this article, I argue that an elevated macro-level perspective is imperative for conducting critical studies in the fields of marketing and consumer research. There are epistemic barriers to operating in this manner, and I offer several suggestions for overcoming these barriers. Finally, I...

  6. How criticality affects student's creativity

    DEFF Research Database (Denmark)

    Adriansen, Hanne Kirstine

    2010-01-01

    In this chapter, I analyse if there is an inherent paradox between creativity and criticality. With critical thinking being among the core values in higher education, can we then also foster creative thinking? In answering this question, I use the masters degree LAICS (Leadership And Innovation...... in Complex Systems) as a case study. Interviews with students are used to shed light on creative teaching and learning. It is shown that creativity can be taught by teaching creatively. I conclude that creativity and criticality are not entirely different ways of thinking and both are important in academia......, but creativity can be hampered by our norms, rules, and structures....

  7. Glucose control in critical care

    Institute of Scientific and Technical Information of China (English)

    2015-01-01

    Glycemic control among critically-ill patients has beena topic of considerable attention for the past 15 years.An initial focus on the potentially deleterious effects ofhyperglycemia led to a series of investigations regardingintensive insulin therapy strategies that targeted tightglycemic control. As knowledge accumulated, the pursuitof tight glycemic control among critically-ill patients cameto be seen as counterproductive, and moderate glycemiccontrol came to dominate as the standard practice inintensive care units. In recent years, there has beenincreased focus on the importance of hypoglycemicepisodes, glycemic variability, and premorbid diabeticstatus as factors that contribute to outcomes amongcritically-ill patients. This review provides a survey ofkey studies on glucose control in critical care, and aimsto deliver perspective regarding glycemic managementamong critically-ill patients.

  8. Mission Critical: Preventing Antibiotic Resistance

    Science.gov (United States)

    ... file Error processing SSI file Mission Critical: Preventing Antibiotic Resistance Recommend on Facebook Tweet Share Compartir Can you ... spp. So, what can we do to prevent antibiotic resistance in healthcare settings? Patients, healthcare providers, healthcare facility ...

  9. Coping with criticism and praise.

    Science.gov (United States)

    Esdale, Lynn; Jahoda, Andrew; Pert, Carol

    2015-05-01

    Through experiencing stigma and discrimination, people with intellectual disability may become more sensitive to criticism from others and be less likely to believe praise. This study compared how people with and without intellectual disability viewed praise and criticism, using a vignette task developed for the study. Participants were asked to imagine someone saying something praiseworthy or critical and were then asked about their emotions, beliefs, and thoughts. People with intellectual disability were more likely to believe and be distressed by criticism. Contrary to predictions, this group were also more likely to believe praise and experience positive affect. The results suggest that the self-perceptions of people with intellectual disabilities is more dynamic and reliant on the views of others.

  10. Hawksbill Sea Turtle Critical Habitat

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These data represent the critical habitat for hawksbill turtle as designated by Federal Register Vol. 63, No. 46701, September 2, 1998, Rules and Regulations....

  11. Leatherback Sea Turtle Critical Habitat

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — These data represent the critical habitat for leatherback turtle as designated by Federal Register Vol. 44, No. 17711, March 23, 1979, Rules and Regulations....

  12. Critical SQG in bounded domains

    OpenAIRE

    Constantin, Peter; Ignatova, Mihaela

    2016-01-01

    We consider the critical dissipative SQG equation in bounded domains, with the square root of the Dirichlet Laplacian dissipation. We prove global a priori interior $C^{\\alpha}$ and Lipschitz bounds for large data.

  13. CRITICAL EVENTS IN CONSTRUCTION PROCESS

    DEFF Research Database (Denmark)

    Jørgensen, Kirsten; Rasmussen, Grane Mikael Gregaard

    2009-01-01

    Function failures, defects and poor communication are major problems in the construction industry. These failures and defects are caused by a row of critical events in the construction process. The purpose of this paper is to define “critical events” in the construction process and to investigate...... cause-effects of failures and defects in the construction industry by using an analytical approach (The bowtie model) which is developed in the accident research. Using this model clarifies the relationships within the chain of failures that causes critical events with undesirable consequences....... In this way the causes of failures and the relationships between various failures are rendered visible. A large construction site was observed from start to finish as the empirical element in the research. The research focuses on all kinds of critical events identified throughout every phase during...

  14. Environmental Education, A Critical Evaluation

    Science.gov (United States)

    Abell, Dana L.

    1970-01-01

    A review of the contents, concept, and furure possibilities of a new journal, with comments on its role in helping generate ideas and encouraging critical thought concerning environmental education. (AL)

  15. Journal of critical globalisation studies

    National Research Council Canada - National Science Library

    The Journal of Critical Globalisation Studies is a peer-reviewed, open-access journal that invites contributors to challenge dominant ideas of globalisation and ideas that globalisation is a singular...

  16. Autoclave nuclear criticality safety analysis

    Energy Technology Data Exchange (ETDEWEB)

    D`Aquila, D.M. [Martin Marietta Energy Systems, Inc., Piketon, OH (United States); Tayloe, R.W. Jr. [Battelle, Columbus, OH (United States)

    1991-12-31

    Steam-heated autoclaves are used in gaseous diffusion uranium enrichment plants to heat large cylinders of UF{sub 6}. Nuclear criticality safety for these autoclaves is evaluated. To enhance criticality safety, systems are incorporated into the design of autoclaves to limit the amount of water present. These safety systems also increase the likelihood that any UF{sub 6} inadvertently released from a cylinder into an autoclave is not released to the environment. Up to 140 pounds of water can be held up in large autoclaves. This mass of water is sufficient to support a nuclear criticality when optimally combined with 125 pounds of UF{sub 6} enriched to 5 percent U{sup 235}. However, water in autoclaves is widely dispersed as condensed droplets and vapor, and is extremely unlikely to form a critical configuration with released UF{sub 6}.

  17. Autism: a "critical period" disorder?

    Science.gov (United States)

    LeBlanc, Jocelyn J; Fagiolini, Michela

    2011-01-01

    Cortical circuits in the brain are refined by experience during critical periods early in postnatal life. Critical periods are regulated by the balance of excitatory and inhibitory (E/I) neurotransmission in the brain during development. There is now increasing evidence of E/I imbalance in autism, a complex genetic neurodevelopmental disorder diagnosed by abnormal socialization, impaired communication, and repetitive behaviors or restricted interests. The underlying cause is still largely unknown and there is no fully effective treatment or cure. We propose that alteration of the expression and/or timing of critical period circuit refinement in primary sensory brain areas may significantly contribute to autistic phenotypes, including cognitive and behavioral impairments. Dissection of the cellular and molecular mechanisms governing well-established critical periods represents a powerful tool to identify new potential therapeutic targets to restore normal plasticity and function in affected neuronal circuits.

  18. Critical Facilities for Coastal Geographies

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The critical facilities data are derived from the USGS Structures Inventory Database (June, 2015). The structures in the derived dataset displays aggregated totals...

  19. Reiki therapy: a nursing intervention for critical care.

    Science.gov (United States)

    Toms, Robin

    2011-01-01

    Complementary and alternative medicine (CAM) is not generally associated with the complexity and intensity of critical care. Most CAM therapies involve slow, calming techniques that seem to be in direct contrast with the fast-paced, highly technical nature of critical care. However, patients in critical care often find themselves coping with the pain and stress of their illness exacerbated by the stress of the critical care environment. Complementary and alternative medicine-related research reveals that complementary therapies, such as Reiki, relieve pain and anxiety and reduce symptoms of stress such as elevated blood pressure and pulse rates. Patients and health care professionals alike have become increasingly interested in complementary and alternative therapies that do not rely on expensive, invasive technology, and are holistic in focus. Reiki is cost-effective, noninvasive, and can easily be incorporated into patient care. The purpose of this article is to examine the science of Reiki therapy and to explore Reiki as a valuable nursing intervention.

  20. Transport signatures of quantum critically in Cr at high pressure.

    Energy Technology Data Exchange (ETDEWEB)

    Jaramillo, R.; Feng, Y.; Wang, J.; Rosenbaum, T. F. (X-Ray Science Division); ( PSC-USR); (Harvard Univ.); (Univ. of Chicago)

    2010-08-03

    The elemental antiferromagnet Cr at high pressure presents a new type of naked quantum critical point that is free of disorder and symmetry-breaking fields. Here we measure magnetotransport in fine detail around the critical pressure, P{sub c} {approx} 10 GPa, in a diamond anvil cell and reveal the role of quantum critical fluctuations at the phase transition. As the magnetism disappears and T {yields} 0, the magntotransport scaling converges to a non-mean-field form that illustrates the reconstruction of the magnetic Fermi surface, and is distinct from the critical scaling measured in chemically disordered Cr:V under pressure. The breakdown of itinerant antiferromagnetism only comes clearly into view in the clean limit, establishing disorder as a relevant variable at a quantum phase transition.

  1. Defining and assessing critical thinking skills for student radiographers

    Energy Technology Data Exchange (ETDEWEB)

    Castle, Alan [University of Portsmouth, School of Health Sciences and Social Work, Centre for Radiography Education, James Watson Building, 2 King Richard 1st Road, Portsmouth PO1 2FR (United Kingdom)], E-mail: alan.castle@port.ac.uk

    2009-02-15

    Developing critical thinking skills is a key aim of higher education and is important in preparing student radiographers for their future careers in clinical practice. The aim of this paper was to attempt to devise and assess six key components of critical thinking appropriate for radiographic practice. Each of the six components was divided into three dimensions and a Critical Thinking Skills Scoring Chart (CTSSC) devised to assess students' written performance against each dimension. Scores revealed that approximately 30% of students were rated as good and approximately 10% of students were rated as poor in each component, although there was some variability between different dimensions. It is suggested that educators need to encourage and support students to develop their critical thinking skills by reviewing their curriculum to clearly define specific skills and ensure that they are appropriately taught and assessed.

  2. Thinking Critically about Critical Thinking: Integrating Online Tools to Promote Critical Thinking

    Directory of Open Access Journals (Sweden)

    B. Jean Mandernach

    2006-01-01

    Full Text Available The value and importance of critical thinking is clearly established; the challenge for instructors lies in successfully promoting students’ critical thinking skills within the confines of a traditional classroom experience. Since instructors are faced with limited student contact time to meet their instructional objectives and facilitate learning, they are often forced to make instructional decisions between content coverage, depth of understanding, and critical analysis of course material. To address this dilemma, it is essential to integrate instructional strategies and techniques that can efficiently and effectively maximize student learning and critical thinking. Modern advances in educational technology have produced a range of online tools to assist instructors in meeting this instructional goal. This review will examine the theoretical foundations of critical thinking in higher education, discuss empirically-based strategies for integrating online instructional supplements to enhance critical thinking, offer techniques for expanding instructional opportunities outside the limitations of traditional class time, and provide practical suggestions for the innovative use of critical thinking strategies via online resources.

  3. Enteral nutrition in critical patients

    OpenAIRE

    Botello Jaimes, Jhon Jairo; Medico Cuidado Intensivo Clínica San Marcel / Jefe de Urgencias y Unidad de Cuidado Especial Clínica San Marcel / Docente Medicina Interna y Cuidado Intensivo Universidad de Manizales / Docente Internado de Urgencias y de PROFUNDIZACION Urgencias, Cuidado Intermedio e Intensivo Universidad de Caldas / Docente del programa de enfermería Universidad Católica de Manizales / Instructor Internacional / Director Fundación Versalles / Candidato a Magíster en Educación Docencia. Envío co rrespondencia:. Avenida Alberto Mendoza No 93-25 - Clínica San Marcel.; González Rincón, Alejandra; Enfermería Universidad Católica de Manizales. Miembro directivo Fundación Versalles. Miembro Correspondiente Asociación Colombiana de Trauma y Miembro Activo de la Sociedad Panamericana de Trauma. Envío correspondencia: alejita_gonzalez12@ hotmail.com . Calle 50 No 24-14 – Fundación Versalles.

    2010-01-01

    Nutritional support in critically ill patients is of vital importance for its impact on morbidity, hospital stay and preservation of tissue mass among others. It must ensure nutritional support to all critically ill patients enterally ideally by reduced bacterial translocationand stimulation of the function of the intestinal villi. In recent years we have studied immunonutrients as glutamine, arginine, fatty acids and nucleotides with promising effects (Immune response, Intestinal barrier or ...

  4. Lecture notes for criticality safety

    Science.gov (United States)

    Fullwood, R.

    1992-03-01

    These lecture notes for criticality safety are prepared for the training of Department of Energy supervisory, project management, and administrative staff. Technical training and basic mathematics are assumed. The notes are designed for a two-day course, taught by two lecturers. Video tapes may be used at the options of the instructors. The notes provide all the materials that are necessary but outside reading will assist in the fullest understanding. The course begins with a nuclear physics overview. The reader is led from the macroscopic world into the microscopic world of atoms and the elementary particles that constitute atoms. The particles, their masses and sizes and properties associated with radioactive decay and fission are introduced along with Einstein's mass-energy equivalence. Radioactive decay, nuclear reactions, radiation penetration, shielding and health-effects are discussed to understand protection in case of a criticality accident. Fission, the fission products, particles and energy released are presented to appreciate the dangers of criticality. Nuclear cross sections are introduced to understand the effectiveness of slow neutrons to produce fission. Chain reactors are presented as an economy; effective use of the neutrons from fission leads to more fission resulting in a power reactor or a criticality excursion. The six-factor formula is presented for managing the neutron budget. This leads to concepts of material and geometric buckling which are used in simple calculations to assure safety from criticality. Experimental measurements and computer code calculations of criticality are discussed. To emphasize the reality, historical criticality accidents are presented in a table with major ones discussed to provide lessons-learned. Finally, standards, NRC guides and regulations, and DOE orders relating to criticality protection are presented.

  5. Teamwork in obstetric critical care

    OpenAIRE

    Guise, Jeanne-Marie; Segel, Sally

    2008-01-01

    Whether seeing a patient in the ambulatory clinic environment, performing a delivery or managing a critically ill patient, obstetric care is a team activity. Failures in teamwork and communication are among the leading causes of adverse obstetric events, accounting for over 70% of sentinel events according to the Joint Commission. Effective, efficient and safe care requires good teamwork. Although nurses, doctors and healthcare staff who work in critical care environments are extremely well t...

  6. Uranium Critical Point Location Problem

    CERN Document Server

    Iosilevskiy, Igor

    2013-01-01

    Significant uncertainty of our present knowledge for uranium critical point parameters is under consideration. Present paper is devoted to comparative analysis of possible resolutions for the problem of uranium critical point location, as well as to discussion of plausible scheme of decisive experiment, which could resolve existing uncertainty. New calculations of gas-liquid coexistence in uranium by modern thermodynamic code are included in the analysis.

  7. Lecture notes for criticality safety

    Energy Technology Data Exchange (ETDEWEB)

    Fullwood, R.

    1992-03-01

    These lecture notes for criticality safety are prepared for the training of Department of Energy supervisory, project management, and administrative staff. Technical training and basic mathematics are assumed. The notes are designed for a two-day course, taught by two lecturers. Video tapes may be used at the options of the instructors. The notes provide all the materials that are necessary but outside reading will assist in the fullest understanding. The course begins with a nuclear physics overview. The reader is led from the macroscopic world into the microscopic world of atoms and the elementary particles that constitute atoms. The particles, their masses and sizes and properties associated with radioactive decay and fission are introduced along with Einstein`s mass-energy equivalence. Radioactive decay, nuclear reactions, radiation penetration, shielding and health-effects are discussed to understand protection in case of a criticality accident. Fission, the fission products, particles and energy released are presented to appreciate the dangers of criticality. Nuclear cross sections are introduced to understand the effectiveness of slow neutrons to produce fission. Chain reactors are presented as an economy; effective use of the neutrons from fission leads to more fission resulting in a power reactor or a criticality excursion. The six-factor formula is presented for managing the neutron budget. This leads to concepts of material and geometric buckling which are used in simple calculations to assure safety from criticality. Experimental measurements and computer code calculations of criticality are discussed. To emphasize the reality, historical criticality accidents are presented in a table with major ones discussed to provide lessons-learned. Finally, standards, NRC guides and regulations, and DOE orders relating to criticality protection are presented.

  8. Lecture notes for criticality safety

    Energy Technology Data Exchange (ETDEWEB)

    Fullwood, R.

    1992-03-01

    These lecture notes for criticality safety are prepared for the training of Department of Energy supervisory, project management, and administrative staff. Technical training and basic mathematics are assumed. The notes are designed for a two-day course, taught by two lecturers. Video tapes may be used at the options of the instructors. The notes provide all the materials that are necessary but outside reading will assist in the fullest understanding. The course begins with a nuclear physics overview. The reader is led from the macroscopic world into the microscopic world of atoms and the elementary particles that constitute atoms. The particles, their masses and sizes and properties associated with radioactive decay and fission are introduced along with Einstein's mass-energy equivalence. Radioactive decay, nuclear reactions, radiation penetration, shielding and health-effects are discussed to understand protection in case of a criticality accident. Fission, the fission products, particles and energy released are presented to appreciate the dangers of criticality. Nuclear cross sections are introduced to understand the effectiveness of slow neutrons to produce fission. Chain reactors are presented as an economy; effective use of the neutrons from fission leads to more fission resulting in a power reactor or a criticality excursion. The six-factor formula is presented for managing the neutron budget. This leads to concepts of material and geometric buckling which are used in simple calculations to assure safety from criticality. Experimental measurements and computer code calculations of criticality are discussed. To emphasize the reality, historical criticality accidents are presented in a table with major ones discussed to provide lessons-learned. Finally, standards, NRC guides and regulations, and DOE orders relating to criticality protection are presented.

  9. Critical health literacy: a review and critical analysis.

    Science.gov (United States)

    Chinn, Deborah

    2011-07-01

    Though there has been a considerable expansion of interest in the health literacy concept worldwide, there has also been criticism that this concept has been poorly defined, that it stretches the idea of "literacy" to an indefensible extent and more specifically, that it adds little to the existing concerns and intervention approaches of the better established discipline of health promotion. This paper takes as a starting point the expanded model of health literacy advanced by Nutbeam (2000) and addresses these concerns by interrogating the concept of "critical health literacy" in order to draw conclusions about its utility for advancing the health of individuals and communities. The constituent domains of critical health literacy are identified; namely information appraisal, understanding the social determinants of health, and collective action, and as far as possible each are clearly delineated, with links to related concepts made explicit. The paper concludes that an appreciation of work undertaken in a range of different disciplines, such as media studies, medical sociology, and evidence-based medicine can enhance our understanding of the critical health literacy construct and help us understand its usefulness as a social asset which helps individuals towards a critical engagement with health information. There is some evidence that aspects of critical health literacy have indeed been found to be a resource for better health outcomes, but more research is needed in this area, both to develop quantitative and qualitative approaches to evaluating health literacy skills, and to offer convincing evidence that investment in programmes designed to enhance critical health literacy are worthwhile.

  10. Dependency in Critically Ill Patients

    Directory of Open Access Journals (Sweden)

    Rumei Yang

    2016-03-01

    Full Text Available By necessity, critically ill patients admitted to intensive care units (ICUs have a high level of dependency, which is linked to a variety of negative feelings, such as powerlessness. However, the term dependency is not well defined in the critically ill patients. The concept of “dependency” in critically ill patients was analyzed using a meta-synthesis approach. An inductive process described by Deborah Finfgeld-Connett was used to analyze the data. Overarching themes emerged that reflected critically ill patients’ experience and meaning of being in dependency were (a antecedents: dependency in critically ill patients was a powerless and vulnerable state, triggered by a life-threatening crisis; (b attributes: the characteristic of losing “self” was featured by dehumanization and disembodiment, which can be alleviated by a “self”-restoring process; and (c outcomes: living with dependency and coping with dependency. The conceptual model explicated here may provide a framework for understanding dependency in critically ill patients.

  11. Dependency in Critically Ill Patients

    Directory of Open Access Journals (Sweden)

    Rumei Yang

    2016-03-01

    Full Text Available By necessity, critically ill patients admitted to intensive care units (ICUs have a high level of dependency, which is linked to a variety of negative feelings, such as powerlessness. However, the term dependency is not well defined in the critically ill patients. The concept of “dependency” in critically ill patients was analyzed using a meta-synthesis approach. An inductive process described by Deborah Finfgeld-Connett was used to analyze the data. Overarching themes emerged that reflected critically ill patients’ experience and meaning of being in dependency were (a antecedents: dependency in critically ill patients was a powerless and vulnerable state, triggered by a life-threatening crisis; (b attributes: the characteristic of losing “self” was featured by dehumanization and disembodiment, which can be alleviated by a “self”-restoring process; and (c outcomes: living with dependency and coping with dependency. The conceptual model explicated here may provide a framework for understanding dependency in critically ill patients.

  12. Empirical Investigation of Critical Transitions in Paleoclimate

    Science.gov (United States)

    Loskutov, E. M.; Mukhin, D.; Gavrilov, A.; Feigin, A.

    2016-12-01

    In this work we apply a new empirical method for the analysis of complex spatially distributed systems to the analysis of paleoclimate data. The method consists of two general parts: (i) revealing the optimal phase-space variables and (ii) construction the empirical prognostic model by observed time series. The method of phase space variables construction based on the data decomposition into nonlinear dynamical modes which was successfully applied to global SST field and allowed clearly separate time scales and reveal climate shift in the observed data interval [1]. The second part, the Bayesian approach to optimal evolution operator reconstruction by time series is based on representation of evolution operator in the form of nonlinear stochastic function represented by artificial neural networks [2,3]. In this work we are focused on the investigation of critical transitions - the abrupt changes in climate dynamics - in match longer time scale process. It is well known that there were number of critical transitions on different time scales in the past. In this work, we demonstrate the first results of applying our empirical methods to analysis of paleoclimate variability. In particular, we discuss the possibility of detecting, identifying and prediction such critical transitions by means of nonlinear empirical modeling using the paleoclimate record time series. The study is supported by Government of Russian Federation (agreement #14.Z50.31.0033 with the Institute of Applied Physics of RAS). 1. Mukhin, D., Gavrilov, A., Feigin, A., Loskutov, E., & Kurths, J. (2015). Principal nonlinear dynamical modes of climate variability. Scientific Reports, 5, 15510. http://doi.org/10.1038/srep155102. Ya. I. Molkov, D. N. Mukhin, E. M. Loskutov, A.M. Feigin, (2012) : Random dynamical models from time series. Phys. Rev. E, Vol. 85, n.3.3. Mukhin, D., Kondrashov, D., Loskutov, E., Gavrilov, A., Feigin, A., & Ghil, M. (2015). Predicting Critical Transitions in ENSO models. Part II

  13. Expressed and perceived criticism, family warmth, and symptoms in schizophrenia.

    Science.gov (United States)

    Medina-Pradas, Cristina; Navarro, J Blas; Pousa, Esther; Montero, M Isabel; Obiols, Jordi E

    2013-01-01

    Criticism directed by caregivers towards a family member with schizophrenia, both from the perspective of the patient and of the caregiver, predicts relapse, although both perspectives differ. This study aims to verify if the same applies to a Mediterranean sample, where criticism is not the main attitude of high expressed emotion families. The Camberwell Family Interview was applied to assess the family's perspective, and the Perceived Criticism and the Family Emotional Involvement and Criticism Scales were used to assess the patients' perspective, in 21 dyads. The association between both perspectives and psychotic symptoms was also examined. Results replicated those of previous studies in other countries, revealing that the perspectives on Criticism of patients and families do not match. The fact that family members also presented positive attitudes towards the patient did not cushion the patient's perceived criticism. Thus, it seems that families considered to be critical may not be perceived as such by the patients. Furthermore, only the warmth from the family's perspective correlated with the symptoms: positive affect proved to be more relevant than negative affect. Therefore, the patients' subjective assessments and family warmth should be included in clinical and research proposals.

  14. Dynamical net-proton fluctuations near a QCD critical point

    CERN Document Server

    Herold, Christoph; Yan, Yupeng; Kobdaj, Chinorat

    2016-01-01

    We investigate the evolution of the net-proton kurtosis and the kurtosis of the chiral order parameter near the critical point in the model of nonequilibrium chiral fluid dynamics. The order parameter is propagated explicitly and coupled to an expanding fluid of quarks and gluons in order to describe the dynamical situation in a heavy-ion collision. We study the critical region near the critical point on the crossover side. There are two sources of fluctuations: non-critical initial event-by-event fluctuations and critical fluctuations. These fluctuations can be distinguished by comparing a mean-field evolution of averaged thermodynamic quantities with the inclusion of fluctuations at the phase transition. We find that while the initial state fluctuations give rise to flat deviations from statistical fluctuations, critical fluctuations reveal a clear structure of the phase transition. The signals of the critical point in the net-proton and sigma field kurtosis are affected by the nonequilibrium dynamics and t...

  15. Fuzzy Critical Path Method Based on Lexicographic Ordering

    Directory of Open Access Journals (Sweden)

    Phani Bushan Rao P

    2012-01-01

    Full Text Available The Critical Path Method (CPM is useful for planning and control of complex projects. The CPM identifies the critical activities in the critical path of an activity network. The successful implementation of CPM requires the availability of clear determined time duration for each activity. However, in practical situations this requirement is usually hard to fulfil since many of activities will be executed for the first time. Hence, there is always uncertainty about the time durations of activities in the network planning.  This has led to the development of fuzzy CPM.  In this paper, we use a Lexicographic ordering method for ranking fuzzy numbers to a critical path method in a fuzzy project network, where the duration time of each activity is represented by a trapezoidal fuzzy number. The proposed method is compared with fuzzy CPM based on different ranking methods of fuzzy numbers. The comparison reveals that the method proposed in this paper is more effective in determining the activity criticalities and finding the critical path.   This new method is simple in calculating fuzzy critical path than many methods proposed so far in literature.  

  16. Quantum-to-classical crossover near quantum critical point.

    Science.gov (United States)

    Vasin, M; Ryzhov, V; Vinokur, V M

    2015-12-21

    A quantum phase transition (QPT) is an inherently dynamic phenomenon. However, while non-dissipative quantum dynamics is described in detail, the question, that is not thoroughly understood is how the omnipresent dissipative processes enter the critical dynamics near a quantum critical point (QCP). Here we report a general approach enabling inclusion of both adiabatic and dissipative processes into the critical dynamics on the same footing. We reveal three distinct critical modes, the adiabatic quantum mode (AQM), the dissipative classical mode [classical critical dynamics mode (CCDM)], and the dissipative quantum critical mode (DQCM). We find that as a result of the transition from the regime dominated by thermal fluctuations to that governed by the quantum ones, the system acquires effective dimension d + zΛ(T), where z is the dynamical exponent, and temperature-depending parameter Λ(T) ∈ [0, 1] decreases with the temperature such that Λ(T = 0) = 1 and Λ(T → ∞) = 0. Our findings lead to a unified picture of quantum critical phenomena including both dissipation- and dissipationless quantum dynamic effects and offer a quantitative description of the quantum-to-classical crossover.

  17. Burnout in critical care nurses: a literature review.

    Science.gov (United States)

    Epp, Kirstin

    2012-01-01

    Burnout and its development in critical care nurses is a concept that has received extensive study, yet remains a problem in Canada and around the world. Critical care nurses are particularly vulnerable to developing burnout due to the chronic occupational stressors they are exposed to, including high patient acuity, high levels of responsibility, working with advanced technology, caring for families in crisis, and involved in morally distressing situations, particularly prolonging life unnecessarily. The purpose of this article is to explore how the chronic stressors that critical care nurses are exposed to contribute to the development of burnout, and strategies for burnout prevention. A review of the literature between the years 2007 and 2012 was conducted and included the search terms burnout, moral distress, compassion fatigue, intensive care, critical care, and nursing. The search was limited to the adult population, English language, and Western cultures. The results revealed that nurse managers play a crucial role in preventing burnout by creating a supportive work environment for critical care nurses. Strategies for nurse managers to accomplish this include being accessible to critical care nurses, fostering collegial relationships among the different disciplines, and making a counsellor or grief team available to facilitate debriefing after stressful situations, such as a death. In addition, critical care nurses can help prevent burnout by being a support system for each other and implementing self-care strategies.

  18. Decision Making and Revealed Preference

    DEFF Research Database (Denmark)

    de la Rosa, Leonidas Enrique

    If our decision-making processes are to some extent shaped by evolutionary pressures and our environment is different from that to which we adapted, some of our choices will not be in our best interest. But revealed preference is the only tool that we have so far to conduct a normative analysis...

  19. Revealed preference with limited consideration

    NARCIS (Netherlands)

    Demuynck, T.; Seel, C.

    2014-01-01

    We derive revealed preference tests for models where individuals use consideration sets to simplify their consumption problem. Our basic test provides necessary and sufficient conditions for consistency of observed choices with the existence of consideration set restrictions. The same conditions can

  20. The archetypal criticism of "Hooshroba Castle" story

    Directory of Open Access Journals (Sweden)

    Jalil Moshydy

    2017-04-01

    Full Text Available Archetypes are some shared concepts which are accumulated in collective unconscious. Archetypal criticism could be defined a branch of literary criticism which deals with analyzing archetypes in a literary work. Archetypes are some concealed things and symbols can make them reveal. Jung believes “archetype’s origin is unknown. They might appear anytime and anywhere; even if transforming of Human origin or racial combinations is ignored by emigration” (Jung, 1352: 106. Archetypal criticism started out of the literature in addition to Jung James Frazer, Scottish anthropologist, the first theorist was that introduced archetypes, but archetypes are known by Jung and mostly consider idiomatic belongs to Jung. Maud Bodkin the first in "archetypal designs in poetry" by interpreting mythic Coleridge's poems, and Eliot, brought literature Jung's ideas and Frazer and others, such as Robert Graves, Robert Graves, Wilson Knight, Richard Chase and … continued Bodkin way to and finally Northrop Frye recognize that the archetypal criticism along with other forms of modern literary criticism and left it of domination anthropology and psychology. Also called the archetypal criticism mythical criticism, because in principle archetypes derived from myths. Myths are related to the course of human civilization be remembered that as human childhood. According to Jungian theory, collective unconscious human thought is consistent with the preliminary stage and belongs to the time until then he was not out to conquer the world and pay attention to the inner the human and tried In mythology says Your inner psychological discoveries pursued. So “The main public symbols inheritable and archetypes perception” (Sattari, 1366: 499. The myth reflects the inner life and the human's wishes that do not go out of her life time and space will not be able to narrow it, that is, why the myths around the world constituent elements are identical or very similar